WO2006068516A1

WO2006068516A1 - Implantable medical devices coated with or containing copper chelating compounds

Info

Publication number: WO2006068516A1
Application number: PCT/NZ2005/000337
Authority: WO
Inventors: Garth James Smith Cooper
Original assignee: Protemix Corporation Limited
Priority date: 2004-12-20
Filing date: 2005-12-20
Publication date: 2006-06-29

Abstract

Medical devices comprising a copper antagonist suitable for introduction into a subject, with or without therapeutic agents.

Description

IMPLANTABLE MEDICAL DEVICES COATED WITH OR CONTAINING COPPER CHELATING COMPOUNDS

FIELD

The subject matter pertains to medical devices, including devices having a composition, surface, or feature that exposes and/or delivers one or more copper antagonists, with or without one or more other therapeutic agents, to tissue and/or fluid with which they come into contact on use.

BACKGROUND

The following description includes information that may be useful in understanding the present invention. It is not an admission that any of the information provided herein is prior art, or relevant, to the presently described or claimed invention, or that any publication or document that is specifically or implicitly identified is prior art or a reference that may be used in evaluating patentability. All documents and other information referred to in this patent are incorporated herein by reference in their entirety.

Over the past two decades, much research effort has been directed towards the development of medical devices and machines that are used in a wide variety of clinical settings to maintain the vital physiological functions of a subject mammal. For example, such devices as catheters, prosthetic heart valves, pacemakers, pulse generators, cardiac defibrillators, arteriovenous shunts, and stents are used extensively in the treatment of cardiac and other diseases. Other examples of medical devices, including screws, anchors, plates, joints and similar devices, for example, are used in orthopaedic surgery. These implantable medical devices are made from a wide variety of materials, including, for example, metals, plastics, and various polymeric materials.

When medical devices, for example, implantable medical devices, are brought into contact with a subject, the subject's natural bodily processes can result in aberrant cellular disposition, growth and/or proliferation, and/or tissue repair. For example, platelets may attach to the medical device which can result in further complications at the site of implantation such as, for example, thrombosis, and/or leukocyte attachment that lead to inflammation, and/or aberrant cellular growth. An example of such a process resulting from the contacting of a medical device with a subject is provided by the use of a stent, which can result in restenosis. Restenosis is thought to involve an overpopulation of smooth muscle cells being deposited on or about the stent, eventually leading to a re-narrowing of the lumen of the blood vessel.

Restenosis, the reclosure of a peripheral or coronary artery following trauma to that artery generally caused by efforts to open a stenosed or occluded portion of the artery, and resulting trauma may be caused by, for example, balloon dilation, ablation, atherectomy or laser treatment of the artery. For example, balloon arterial injury reportedly results in endothelial denudation and subsequent regrowth of dysfunctional endothelium (Saville, Analyst, 83 :670-672, 1958) that may contribute to the local smooth muscle cell proliferation and extracellular matrix production that results in reocclusion of the arterial lumen. Restenosis typically occurs in approximately 10-50% of patients undergoing such angioplasty procedures. Restenosis is believed to be a natural healing process in reaction to the injury of the arterior wall caused by such angioplasty procedures. The healing process begins with the thrombotic mechanism at the site of the injury. The final steps of the healing process can be intimal hyperplasia, the uncontrolled migration and proliferation of medial smooth muscle cells, combined with their extracellular matrix production, until the artery is again stenosed or occluded.

There is a need for medical devices that will aid in ameliorating tissue damage and/or enhancing tissue repair and/or limiting or inhibiting complications associated with using or implanting medical devices in a subject, such as for example, restenosis and attachment and/or growth or proliferation of cells, particularly attachment and/or growth or proliferation of platelets, leukocytes and similar cells, etc. It would be desirable to provide a medical device that aids in prevention, amelioration or treatment of damaged tissue and/or enhancement of tissue repair and/or inflammation. SUMMARY

The inventions described and claimed herein have many attributes and embodiments including, but not limited to, those set forth or described or referenced in this Summary. The inventions described and claimed herein are not limited to or by the features or embodiments identified in this Summary, which is included for purposes of illustration only and not restriction.

The present invention is in part directed to new medical devices which may be used, for example, in treating and preventing various diseases, disorders and/or conditions, including excess copper related diseases, disorders, and/or conditions, or those related to unwanted copper whether or not in excess, in a mammal including, but not limited to, the kind referenced herein, and/or enhancing tissue repair processes and/or ameliorating tissue damage and/or inflammation.

In one aspect, the invention relates a medical device for use or implantation in a subject, wherein said medical device comprises a copper antagonist.

In one aspect, the invention relates to an implantable medical device comprising a copper antagonist releasable upon insertion of the medical device to or within a subject

In one aspect of the invention, medical devices comprising a copper antagonist, may include, for example, stents, balloons, prosthetic heart valves, annuloplasty rings, ventricular assist devices, including left ventricular assist devices, right ventricular assist devices, and biventricular assist devices, grafts, shunts, sewing rings (including those having silicone or polyurethane inserts), polyester fabric encasements, medical leads, orthopedic plates, bone pins, bone substitutes, anchors, joints, screws, ophthalmic implants (including, for example, orbital implants, lens implants, corneal implants (including intrasomal corneal ring segments (INTACS)), and microchips), catheters, cannulae, pulse generators, cardiac defibrillators, arteriovenous shunts, pacemakers, sutures, suture anchors, staples, anastomosis devices, vertebral disks, hemostatic barriers, clamps, clips, vascular implants, tissue adhesives and sealants, tissue scaffolds, intraluminal devices, and vascular supports. In one aspect of the invention, the medical device comprising a copper antagonist is a ventricular assist device. In another aspect of the invention, the medical device comprising a copper antagonist is a left ventricular assist device. In yet another aspect of the invention, the medical device comprising a copper antagonist is a right ventricular assist device.

In one aspect of the invention, the medical device comprising a copper antagonist comprises a stent. In another aspect, the stent comprising a copper antagonist is a drug-eluting stent.

In another aspect of the invention, a medical device comprises an instrument, such as a catheter, for example, or an implant suitable for introduction into a subject of which at least a portion comprises a copper antagonist that is available to chelate or bind copper in the subject when the device is used.

In one embodiment, the release rate of the copper antagonist is controlled. In one embodiment of the invention, the copper antagonist may be released slowly or over a sustained period. In one embodiment the copper antagonist is released, for example, over the period of the resorption or degradation of the body of the medical device, or a portion thereof.

In one embodiment of the invention, the surface of the medical device comprises a copper antagonist. In one aspect of the invention a copper antagonist is bound, either directly or indirectly, to a medical device. In another aspect of the invention, a copper antagonist is bound, directly or indirectly, to a surface of a medical device. In another aspect of the invention, this surface contacts a tissue within a subject. In one embodiment, the target tissue is heart tissue or vascular tissue. In yet another embodiment, this surface contacts a site of injury or potential injury. In another embodiment, the medical device provides for surface contact release of the copper antagonist.

In one embodiment, a copper antagonist is present in a coating on a surface of the medical device. Such coatings include, for example, synthetic or natural matrices, for example, fibrin or acetate-based polymers, mixtures of polymers or copolymers, which can also be bioresorbable or biodegradable matrices, and which matrices have or include or incorporate a copper antagonist. Such matrices can, for example, provide for surface contact or metered or sustained release of one or more copper antagonists.

In another embodiment, the device comprising a copper antagonist may be formed, at least in part, for example, from a biodegradable or bioresorbable polymer material. Polymer materials can include, for example, but are not limited to, nylon, polyethylene perthalate, polytetrafluoroethylene, etc. Other polymers may also include, for example, hexafluoropropylene (HFP), tetrafluoroethylene (TFE), vinylidenefluoride, 1-hydropentafluoropropylene, perfluoro(methyl vinyl ether), chlorotrifluoroethylene (CTFE), pentafluoropropene, trifluoroethylene, hexafluoroacetone and hexafluoroisobutylene.

In another aspect, the surface of a medical device comprising a copper antagonist may be composed of organic materials or a composite of organic and inorganic materials. Examples of such materials include, but are not limited to, for example, synthetic polymers or copolymers containing one or more copper antagonists, surfaces upon which a functionalized monolayer containing the copper antagonist is adsorbed or otherwise attached, or synthetic polymeric materials or proteins blended with one or more copper antagonists. In another aspect, all or a portion of the medical device is coated with a copper antagonist, either as the coating per se or in a coating matrix, for example; or all or a portion of the medical device may be produced from a material which includes a copper antagonist, for example, a polymer which has admixed therewith a copper antagonist or which includes a functionalized copper antagonist; or all or a portion of the tissue-contacting surfaces of the medical device may be derivatized with a copper antagonist.

The medical device can be coated using any one or more methods known in the art, for example, dip coating, spray coating, sponging or brushing. The coating may contain the copper antagonist in a weight percentage range of from about 0.0001% to about 30%, for example, although other amounts are contemplated and may be used. Thus, according to one option, the coating may contain copper antagonist in a weight percentage range of about 0.001% to about 25%, alternatively in a range of about 0.01% to about 20%, about 0.1% to about 15%, about 0.5% to about 12%, about 1% to about 10%, about 2% to about 10%, about 5% to about 10%, about 0.01% to about 5%, about 0.1% to about 5% or about 0.5% to about 5%. The weight percentage for the copper antagonist will be adjusted as appropriate, in view of considerations which include, but are not limited to, the following: the dose of copper antagonist to be delivered locally, the rate of release of copper antagonist from the coating and the time period for delivery of copper antagonist. Included within the coating may be further suitable excipients, for example, a polymer and/or those excipients that aid in the binding of the coating to the copper antagonist (or visa versa) and/or that aid in the release of the copper antagonist. Alternatively, the excipients can be bound to the copper antagonist, for example, a polymer or other substance that entraps the copper antagonist on or within the surface of the medical device. In another aspect, all or a portion or portions of the medical device may be coated with the copper antagonist per se or with a pharmaceutically acceptable carrier or excipient comprising the copper antagonist which serves as a coat or coating matrix. This may be a solid, liquid, gel or semisolid consistency, for example. The carrier or matrix can be made of or include agents which provide, for example, for metered or slow or sustained release of the copper antagonist and/or other therapeutic agent(s). The coating, for example, can include albumin that can be either, for example, human or bovine, including humanized bovine serum albumin. A coating may be applied as a single coating or in multiple coatings or layers. The multiple coatings or layers can include varying ratios of copper antagonist-to-carrier to vary the release rate of the drug over time. The multiple coatings or layers can also include different medicaments in accordance with a desired treatment plan. In one embodiment, the coating comprises a plurality of coatings or layers of a polymer/copper antagonist mixture applied to the medical device. In another embodiment, the copper antagonist-to-polymer ratio in the plurality of coatings or layers varies.

In another aspect, copper antagonists may be impregnated or otherwise incorporated into the body of all or a portion of the medical device. Additionally, the copper antagonist may be provided in one or more reservoirs or channels formed in the medical device, optionally with a coating or membrane of biocompatible material applied over the medical device to control diffusion of the drug from the reservoirs/channels to the tissue. In one embodiment, the medical device comprises at least one channel formed in an outer surface thereof, and wherein the copper antagonist is included on and/or within at least one channel.

In another aspect, if a "burst effect" is desired, the copper antagonist may be applied at the outer layer of the medical device (and/or within the device, as well) so that an initial amount of the copper antagonist is promptly released when it comes into contact with tissue. Remaining amounts, if any, of copper antagonist included in the inner layers will be released over time as the copper antagonist diffuses through the material. The copper antagonist may be provided either in liquid or solid form.

In another aspect, the coating applied to the medical device can be "recharged", for example, by way of a catheter or other tubing capable of infusing a copper antagonist donor to a previously coated surface or impregnated or other device comprising a copper antagonist. For example, the copper antagonist may be functionalized with, for example, sulphur to form an S- functionalized-copper antagonist-protein which will lose potency in vivo as the S-functionalized-copper antagonist-protein is metabolized, leaving un- derivatized protein. The surface coating can be "recharged" by infusing a copper antagonist or copper antagonist donor capable of binding the un-derivatized protein.

In another aspect of the invention, the derivatization of an artificial surface with a copper antagonist provides for the amelioration of tissue damage and/or enhancement of tissue repair, such as, for example, the prevention of the deposition of platelets and for preventing thrombus formation on the artificial surface, as well as the prevention and/or amelioration of imflammation. The artificial surfaces may be composed of organic materials or a composite of organic and inorganic materials. Examples of such materials include synthetic polymers or copolymers containing one or more copper antagonists, surfaces upon which a functionalized monolayer containing the copper antagonist is absorbed, or synthetic polymeric materials or proteins which are blended with the copper antagonist.

In one embodiment of the invention, the localized and/or time-related presence of a copper antagonist administered in a physiologically effective form is efficacious in diminishing, deterring or preventing vascular damage, including inflammation, after or as a result of instrumental intervention, such as angioplasty, catheterization, or the introduction of a stent {e.g., a Palmaz-Schatz stent) or other surgical or indwelling medical device. Local administration of a stable copper antagonist inhibits neointimal proliferation and platelet deposition following vascular arterial balloon injury, for example. This strategy for the local delivery of a long-lived copper antagonist is useful, among other things, for the treatment of vascular injury following angioplasty.

In a further embodiment, the invention provides for the localized use of a functionalized copper antagonist-protein, particularly those which do not elicit any significant or otherwise undesired immune response. Such functionalized copper antagonist-proteins, such as, for example, functionalized copper antagonist-albumins, can be present as polymeric chains or three dimensional aggregates where the functionalized copper antagonist-protein is the monomeric unit. The protein of the monomeric unit can be a functional subunit of full-length native protein or can be a protein to which has been attached an additional moiety, such as a polypeptide, which can aid, for example, in localization. The aggregates may be multiple inter-adherent monomeric units which can optionally be linked by disulfide bridges. Additionally, devices which have been substituted or coated with one or more functionalized copper antagonist-proteins may be dried and stored. In another aspect, the invention relates to a method of preventing and/or treating damage associated with the use or implantation of a medical device in a subject comprising introducing into said subject a medical device of which at least a portion comprises a copper antagonist, wherein said damage is prevented, ameliorated and/or delayed.

In another aspect, the invention related to a method of preventing and/or treating damage associated with the use or implantation of a medical device in a subject comprising use or implantation of a medical device which comprises a copper antagonist that is releasable at its point of contact, wherein the damage is prevented, amerliorated and/or delayed.

In yet another aspect, the invention relates to a method of preventing and/or treating adverse effects associated with the use or insertion of a medical device in a subject, wherein a copper antagonist is locally administered at the site of contact of said medical device, before and/or during said use or insertion, wherein said adverse effects are prevented, ameliorated or delayed.

In another aspect, the invention relates to a method of treating a damaged vessel in a subject in need thereof which comprises introducing into said vessel at the site of damage a catheter, a balloon, a graft, a stent or a shunt comprising a copper antagonist. In another aspect, the invention comprises a method of ameliorating tissue damage and/or enhancing tissue repair by locally administering one or more copper antagonists to the site of tissue in need thereof. Such tissue may be damaged, or have been damaged, for example, as a result of the use of a medical device in an invasive procedure. Thus, for example, in treating blocked vasculature, by, for example, angioplasty, damage to the blood vessel can result. Additionally, tissue damage can result when medical devices are left within a subject for an extended period of time. The aforementioned damage may be treated by use of a copper antagonist. In addition to ameliorating tissue damage and/or enhancing repair of the damaged tissue, such treatment can also be used to prevent and/or alleviate and/or delay occlusions, including for example, acute occlusion and reocclusions. Treatment may also be used to prevent and/or ameliorate and/or delay thrombosis and restenosis. Treatment may also be used to prevent and/or ameliorate and/or delay inflammation. Treatment may also be used to prevent and/or ameliorate and/or delay oxidative damage, including, for example oxidative damage by free radicals, including superoxide damage. In other aspect, the invention, treatment may also be used to treat and/or prevent and/or alleviate and/or delay atherosclerotic lesions, undesired cell proliferation and migration, undesired immune responses, and fibrosis.

In one embodiment the invention comprises a method of ameliorating tissue damage and/or enhancing tissue repair wherein the subject is a mammal. In another embodiment the invention comprises a method of ameliorating tissue damage and/or enhancing tissue repair wherein the subject is human. In another embodiment the invention comprises a method of ameliorating tissue damage and/or enhancing tissue repair wherein the subject is selected from the group consisting of domestic and pet animals (for example, horses, dogs and cats), sports animals (for example, horses and dogs), farm animals, and zoo animals.

In one embodiment the medical device comprising a copper antagonist can include one or more other therapeutic agents. In one embodiment, the therapeutic agents may be applied or included directly with the copper antagonist. In another embodiment, the therapeutic agent may be applied or included in the same coating or coating layer with the copper antagonist. In yet another embodiment, the therapeutic agent may be applied or included in separate coating or coating layer or a separate portion of the device. Therapeutic agents may include, for example, anti-thrombogenic agents, anticoagulants, antiplatelet agents, thrombolytics, antiproliferatives, anti-inflammatory agents, statins, α-adrenergic receptor antagonists, βrselective adrenergic antagonists, ACE inhibitors, calcium channel blockers, angiotensin II receptor antagonists, vasodilators, anti-proliferative/antimitotic agents, immunosuppressive agent, agents that inhibit hyperplasia and in particular restenosis, smooth muscle cell inhibitors, antibiotics, growth factors, growth factor inhibitors, cell adhesion inhibitors, cell adhesion promoters and drugs that may enhance the formation of healthy neointimal tissue, including endothelial cell regeneration, etc. Anti-thrombogenic agents, may include, for example, heparin, warfarin, hirudin and its analogs, aspirin, indomethacin, dipyridamole, prostacyclin, prostaglandin E, sulfinpyrazone, abciximab, eptifabatide, phenothiazines (such as chlorpromazine or trifluperazine) RGD (arginine-glycine-aspartic acid) peptide or RGD peptide mimetics, agents that block platelet glycoprotein Hb-IIIa receptors (such as C-7E3), ticlopidine or the thienopyridine known as clopidogrel.

Statins, may include, for example, simvastatin, atorvastatin, lovastatin, pravastatin, and fluvastatin. α-adrenergic receptor antagonists may include, for example, prazosin, terazosin, doxazosin, ketanserin, indoramin, urapidil, clonideine, guanabenz, guanfacine, guanadrel, reserpine, and metyrosine. β^selective adrenergic antagonists may include, for example, metoprolol, atenolol, esmolol, acebutolol, bopindolol, carteolol, oxprenolol, penbutolol, medroxalol, bucindolol, levobunolol, metipranolol, bisoprolol, nebivolol, betaxolol, celiprolol, sotalol, propafenone, propranolol, timolol maleate, and nadolol.

ACE inhibitors may include, for example, captopriol, fentiapril, pivalopril, zofenopril, alacepril, enalapril, enalaprilat, enalaprilo, lisinopril, benazepril, quinapril, moexipril.

Calcium channel blockers may include, for example, nisoldipine, verapamil, diltiazem, nifedipine, nimodipine, felodipine, nicardipine, isradipine, amlodipine, and bepridil.

Angiotensin II receptor antagonists may include, for example, losartan, candesartan, irbesartan, valsartan, telmisartan, eprosartan, and olmesartan medoxomil.

Vasodilators may include, for example, hydralazine, minoxidil, sodium nitroprusside, diazoxide, bosentan, eporprostenol, treprostinil, and iloprost*

Anti-inflammatory agents may include, for example, steroids (including, for example, Cortisol, cortisone, fludrocortisone, prednisone, prednisolone, 6-α- methylprednisolone, triamcinolone, betamethasone, corticosterone, budesonide, estrogen, sulfasalazine, mesalamine and dexamethasone), non-steroidal agents

(including, for example, salicylic acid derivatives i.e. aspirin; para-aminophenol derivatives e.g. acetaminophen; indole and indene acetic acids, e.g.indomethacin, sulindac, and etodalac; heteroaryl acetic acids e.g. tolmetin, diclofenac, and ketorolac; arylpropionic acids e.gdbuprofen and derivatives; anthranilic acids e.g.mefenamic acid, and meclofenamic acid; enolic acids e.g.piroxicam, tenoxicam, phenylbutazone, and oxyphenthatrazone); and nabumetone.

Inmnnosuppresant agents may include, for example, sirolimus, cyclosporine, tacrolimus, azathioprine, and mycophenolate mofetil. Anti-proliferative/antimitotic agents may also be used as therapeutic agents, including, for example, such as vinca alkaloids (e.g. vinblastine, vincristine, and vinorelbine), paclitaxel, epidipodophyllotoxins (e.g. etoposide, teniposide), antibiotics (dactinomycin (actinomycin D), daunorubicin, doxorubicin and idarubicin), anthracyclines, mitoxantrone, bleomycins, plicamycin (mithramycin) and mitomycin.

Antiplatelet agents including, for example, acetylsalicylic acid, dipyridamole, clopidogrel, ticlopidine, abciximab, eptifbatide, tirofiban, reversable COX-I inhibitors, BPIIIb/IIIa blockers, TP antagonists, and P2Y12 antagonists. In one embodiment, the copper antagonist is a copper chelator. In another embodiment, the copper antagonist chelates copper (II).

Preferably, the copper antagonist binds or chelates copper (II). Additional copper antagonist compounds, including acyclic and cyclic compounds, are provided below in reference to Formula I and Formula II as described herein, and salts thereof. Compounds suitable as copper antagonists include cyclic and acyclic compounds according to Formula I:

FORMULA I wherein X₁, X₂, X₃ and X₄ are independently selected from the group consisting of N, S and O; R₁ R₂ R₃ R₄ R₅ and R₆ are independently selected from the group consisting of H, Cl to ClO straight chain or branched alkyl, C3 to ClO cycloalkyl, Cl to C6 alkyl C3 to ClO cycloalkyl, anyl, anyl substituted with 1 to 5 substituents, heteroaryl, fused aryl, Cl to C6 alkyl aryl, Cl to C6 alkyl aryl substituted with 1 to 5 substituents, Cl to C5 alkyl heteroaryl, Cl to C6 alkyl fused aryl, -CH₂COOH, -CH₂SO₃H, -CH₂PO(OH)₂, and -CH₂P(CH₃)O(OH); nl, n2 and n3 are independently 2 or 3 and each of R_7, Rg_, R₉₎ Ri_0, R₁₁ and R₁₂ is independently selected and is selected from the group consisting of H, Cl to ClO straight chain or branched alkyl, C3 to ClO cycloalkyl, Cl to C6 alkyl, C3 to ClO cycloalkyl, aryl, aryl substituted with 1 to 5 substituents, heteroaryl fused aryl, Cl to C6 alkyl aryl, Cl to C6 alkyl aryl substituted with 1 to 5 substituents, Cl to C5 alkyl heteroaryl, Cl to C6 fused aryl, provided that when X₁ is S or O, then R₂ is absent; when X₂ is S or O, then R₃ is absent, when X₃ is S or O, then R₄ is absent and when X₄ is S or O, then R₅ is absent.

Optionally, one or more of Ri₁ R_2, R_3j R_4, R₅ and R₆ may be functionalized for attachment to groups which include, but are not limited to peptides, proteins, polyethylene glycols (PEGs) and other suitable chemical entities in order to modify the overall pharmacokinetics, deliverability and/or half lives of the constructs. Examples of such functionalization include, but are not limited to, Cl to ClO alkyl-CO-peptide, Cl to ClO alkyl-CO-protein, Cl to ClO alkyl-CO-PEG, Cl to ClO alkyl-NH-peptide, Cl to ClO alkyl-NH-protein, Cl to ClO alkyl-NH-CO-PEG, Cl to ClO alkyl-S-peptide, and Cl to ClO alkyl-S-protein.

In addition, optionally are one or more Of R₇, R₈, R₉, R_1O, R₁₁ ^and Ri 2 may be functionalized for attachment to groups which include, but are not limited to, peptides, proteins, polyethylene glycols (PEGs) and other suitable chemical entities in order to modify the overall pharmacokinetics, deliverability and/or half lives of the constructs. Examples of such functionalization include, but are not limited to, Cl to ClO alkyl-CO-peptide, Cl to ClO alkyl-CO-protein, Cl to ClO alkyl-CO-PEG, Cl to ClO alkyl-NH-peptide, Cl to ClO alkyl-NH-protein, Cl to ClO alkyl-NH-CO-PEG, Cl to ClO alkyl-S-peptide and Cl to ClO alkyl-S-protein.

One group of suitable compounds of Formula I include those wherein Ri, R₂, R_3, R_4, R₅ and R₆ are independently selected from H, Cl to C6 alkyl, -CH₂COOH, -CH₂SO₃H, -CH₂PO(OH)₂ and -CH₂P(CH₃)O(OH); and each R₇, R_8, R_9, Ri_0, R₁₁ and Ri₂ is independently selected from H and Cl to C6 alkyl. In one aspect, suitable compounds include those wherein at least one of R₁ and R₂ and at least one of R₅ and R₆ is H or Cl to C6 alkyl. According to this aspect, suitably R₃ and R₄ are selected from H or Cl to C6 alkyl; more particularly, R_1, R_2; R_5> and R₆ are selected from H or Cl to C6 alkyl. One sub-group of suitable compounds include those wherein X₂ and X₃ are N and nl, n2 and n3 are 2, or nl and n3 are 2 and n2 is 3. In this sub-group, Ri_, R₆₁ R_7; R_8; R₉₎ Ri_0, Rn_, and R₁₂ are independently selected from H and Cl to C3 alkyl. According to another subgroup of suitable compounds, all of X_1; X_2, X_3, and X₄ are suitably N or, alternatively, one OfX₁ and X₄ is S and X₂ and X₃ are N or S. Suitable copper antagonist compounds of Formula I include, for example:

SH-CH₂-CH₂-NH-CH₂-CH₂-NH-CH₂-CH₂-NH₂, SH-CH₂-CH₂-S-CH₂-CH₂-NH-CH₂-CH₂-NH₂, NH₂-CH₂-CH₂-NH-CH₂-CH₂-S-CH₂-CH₂-SH,' NH₂-CH₂-CH₂-S-CH₂-CH₂-S-CH₂-CH₂-SH, SH-CH₂-CH₂-S-CH₂-CH₂-S-CH₂-CH₂-SH,

NH₂-CH₂-CH₂-NH-CH₂-CH₂-CH₂-NH-CH₂-CH₂-NH₂, NZ2005/000337

15 SH-CH₂-CH₂-NH-CH₂-CH₂-CH₂-NH-CH₂-CH₂-NH₂,

SH-CH₂-CH₂-S-CH₂-CH₂-CH₂-NH-CH₂-CH₂-NH₂, NH₂-CH₂-CH₂-NH-CH₂-CH₂-CH₂-S-CH₂-CH₂-SH, NH₂-CH₂-CH₂-S-CH₂-CH₂-CH₂-S-CH₂-CH₂-SH, and SH-CH₂-CH₂-S-CH₂-CH₂-CH₂-S-CH₂-CH₂-SH.

Additional compounds suitable as copper antagonists include cyclic and acyclic compounds according to Formula II:

FORMULA II wherein X₁, X₂ and X₃ are independently selected from the group consisting of N, S and O; Ri₅ R_2j R_3; R₅ and R₆ are independently selected from the group consisting of H, C₁ to Ci₀ straight chain or branched alkyl, C3 to ClO cycloalkyl, Cl to C6 alkyl C3 to ClO cycloalkyl, anyl, anyl substituted with 1 to 5 substituents, heteroaryl, fused aryl, Cl to C6 alkyl aryl, Cl to C6 alkyl aryl substituted with 1 to 5 substituents, Cl to C5 alkyl heteroaryl, Cl to C6 alkyl fused aryl, -CH₂COOH, -CH₂SO₃H, -CH₂PO(OH)₂, and -CH₂P(CH₃)O(OH); nl and n2 are independently 2 or 3 and each of R_7s R_8; R₉ and R_10, is independently selected and is selected from the group consisting of H, Cl to ClO straight chain or branched alkyl, C3 to ClO cycloalkyl, Cl to C6 alkyl, C3 to ClO cycloalkyl, aryl, aryl substituted with 1 to 5 substituents, heteroaryl fused aryl, Cl to C6 alkyl aryl, Cl to C6 alkyl aryl substituted with 1 to 5 substituents, Cl to C5 alkyl heteroaryl, Cl to C6 fused aryl, provided that when X₁ is S or O, then R₂ is absent; when X₂ is S or O, then R₃ is absent, and when X₃ is S or O, then R₅ is absent.

Optionally, one or more of R₁ R₂ R₃ R₅ and R₆ may be functionalized for attachment to groups which include, but are not limited to peptides, proteins, polyethylene glycols (PEGs) and other suitable chemical entities in order to modify the overall pharmacokinetics, deliverability and/or half lives of the constructs. Examples of such functionalization include, but are not limited to, Cl to ClO alkyl-CO-peptide, Cl to ClO alkyl-CO-protein, Cl to ClO alkyl-CO-PEG, Cl to ClO alkyl-NH-peptide, Cl to ClO alkyl-NH-protein, Cl to ClO alkyl-NH-CO-PEG, Cl to ClO alkyl-S-peptide, and Cl to ClO alkyl-S-protein.

In addition, optionally are one or more of R₇, R₈, R₉, and R_1O may be functionalized for attachment to groups which include, but are not limited to, peptides, proteins, polyethylene glycols (PEGs) and other suitable chemical entities in order to modify the overall pharmacokinetics, deliverability and/or half lives of the constructs. Examples of such functionalization include, but are not limited to, Cl to ClO alkyl-CO-peptide, Cl to ClO alkyl-CO-protein, Cl to ClO alkyl-CO-PEG, Cl to ClO alkyl-NH-peptide, Cl to ClO alkyl-NH-protein, Cl to ClO alkyl-NH-CO-PEG, Cl to ClO alkyl-S-peptide and Cl to ClO alkyl-S-protein.

One group of suitable compounds of Formula II include those wherein Ri, R₂, R₃, R₅ and R₆ are independently selected from H, Cl to C6 alkyl, -CH₂COOH, -CH₂SO₃H, -CH₂PO(OH)₂ and -CH₂P(CH₃)O(OH); and each R_7, R_8, R₉ and Rio is independently selected from H and Cl to C6 alkyl. In one aspect, suitable compounds include those wherein at least one of Ri and R₂ and at least one Of R₅ and R₆ is H or Cl to C6 alkyl. According to this aspect, suitably R₃ is selected from H or Cl to C6 alkyl; more particularly, Ri₅ R_2j R_5; and R₆ are selected from H or Cl to C6 alkyl. One sub-group of suitable compounds include those wherein Ri_, R₆₅ R_7; R₈ R₉ and Ri_0, are independently selected from H and Cl to C3 alkyl. According to another sub-group of suitable compounds, all of Xi_, X₂ and X₃ are suitably N or, alternatively, one of Xi and X₃ is S and X₂ is N or S.

In another embodiment, the copper antagonist is a copper-chelating tetramine, for example, including 2,3,2 tetramine, 2,2,2 tetramine, and 3,3,3 tetramine. Other copper antagonist compounds are triethylenetetramine, triethylenetetramine acid addition salts, and triethylenetetramine active metabolites including, for example, N-acetyl triethylenetetramine, and analogues, derivatives, and prodrugs of any of the foregoing, and functionalized compounds. In one embodiment, triethylenetetramine is rendered less basic (for example, as an acid addition salt). Salts of triethylenetetramine (which optionally can be salts of a prodrug of triethylenetetramine or a copper chelating metabolite of triethylenetetramine) include, in one embodiment, acid addition salts such as, for example, those of suitable mineral or organic acids. Salts of triethylenetetramine (such as acid addition salts, of acids, including hydrochloric, succinic, maleic, and fumaric acids, e.g., triethylenetetramine dihydrochloride, or triethylenetetramine disuccinate, triethylenetetramine tetramaleate, triethylenetetramine tetrafumarate or other acceptable hydrochloride, succinate, maleate or fumarate salts) act as copper antagonists, e.g., as copper-chelating agents, that aid in the elimination of copper from the body by forming a stable soluble complex that is readily excreted by the kidney.

In another embodiment, triethylenetetramine is modified, i.e., it may be as an analogue or derivative of triethylenetetramine (or an analogue or derivative of a copper-chelating metabolite of triethylenetetramine, for example, N-acetyl triethylenetetramine). Derivatives of triethylenetetramine or triethylenetetramine salts or analogues include those modified with polyethylene glycol (PEG). Analogues of triethylenetetramine include, for example, compounds in which one or more sulfur molecules is substituted for one or more of the NH groups in triethylenetetramine. Other analogues include, for example, compounds in which triethylenetetramine has been modified to include one or more additional -CH₂ groups. One or more hydroxyl groups may also be substituted for one or more amine groups to create an analogue of triethylenetetramine (with or without the substitution of one or more sulfurs for one or more nitrogens).

In another embodiment, triethylenetetramine is delivered as a prodrug of triethylenetetramine or a copper chelating metabolite of triethylenetetramine. In another embodiment the copper chelator is a triethylenetetramine active agent. Triethylenetetramine active agents include, for example, triethylenetetramine, salt(s) of triethylenetetramine, a triethylenetetramine prodrug or a salt of such a prodrug, a triethylenetetramine analogue or a salt or prodrug of such an analogue, and/or at least one active metabolite of triethylenetetramine or a salt or prodrug of such a metabolite, including but not limited to N-acetyl triethylenetetramine and salts and prodrugs of N-acetyl triethylenetetramine. Triethylenetetramine active agents also include the analogues of Formula I and II and/or prodrugs and/or salts of said prodrugs of said analogues.

DETAILED DESCRIPTION As used herein, a "copper antagonist" is a pharmaceutically acceptable compound that binds or chelates copper, preferably copper (II), in vivo for removal. Copper chelators are presently preferred copper antagonists. Copper (II) chelators, and copper (II) specific chelators (i.e., those that preferentially bind copper (II) over other forms of copper such as copper (I)), are especially preferred. "Copper (II)" refers to the oxidized (or +2) form of copper, also sometimes referred to as Cu⁺².

The term "damage associated with the use or implantation of a medical device" refers to damage or injury to tissue resulting from the insertion, presence or removal of a medical device and is evidenced by one or more conditions which include an inflammatory response, a proliferative response including neointimal proliferation, cellular proliferation, removal of endothelium and damage to smooth muscle cells, stenosis following angioplasty or insertion and removal of a medical device, neointimal hyperplasia leading to stenosis, platelet adhesion restenosis, and other inflammatory processes which are associated with implantation, use and removal of a medical device.

As used herein, a "disorder" is any disorder, disease, or condition that would benefit from an agent that reduces local or systemic copper or copper concentrations. Particularly preferred are agents that reduce extracellular copper or extracellular copper concentrations (local or systemic) and, more particularly, agents that reduce extracellular copper (generally, copper (H)) or extracellular copper (II) concentrations (local or systemic). Disorders include, but are not limited to, damage to tissue (including, e.g., heart, liver, kidney, brain tissue) and vascular damage.

As used herein, "mammal" refers to any animal classified as a mammal, including humans, domestic and farm animals, and zoo, sports, or pet animals, such as dogs, horses, cats, sheep, pigs, cows, etc. The preferred mammal herein is a human.

As used herein, "pharmaceutically acceptable salts" refers to salts prepared from pharmaceutically acceptable non-toxic bases or acids including inorganic or organic bases and inorganic or organic acids the like. When the copper antagonist compound is basic, salts may be prepared from pharmaceutically acceptable non-toxic acids, including inorganic and organic acids. Such acids include acetic, benzenesulfonic, benzoic, camphorsulfonic, citric, ethanesulfonic, fumaric, gluconic, glutamic, hydrobromic, hydrochloric, isethionic, lactic, maleic, malic, mandelic, methanesulfonic, mucic, nitric, pamoic, pantothenic, phosphoric, succinic, sulfuric, tartaric, p-toluenesulfonic acid, and the like. Particularly preferred are hydrochloric and succinic acid salts, as well as fumaric and maleic salts. Most preferred are dihydrochloride, disuccinate, tetramaleate and tetrafumarate salts.

As used herein, "preventing" means preventing in whole or in part, or ameliorating or controlling.

As used herein, a "therapeutically-effective amount" or a "pharmaceutically-effective amount" in reference to the compounds or compositions of the instant invention refers to the amount sufficient to induce a desired biological result. That result can be alleviation of the signs, symptoms, or causes of a disease or disorder or condition, or any other desired alteration of a biological system. In the present invention, the result will typically involve the prevention, decrease, or reversal of tissue injury or damage, including inflammation, in whole or in part. >

As used herein, the term "treating" refers to both therapeutic treatment and prophylactic or preventative measures. Those in need of treatment include those already with the disorder as well as those prone to having the disorder or diagnosed with the disorder or those in which the disorder is to be prevented.

The term "medical device" refers to an instrument, apparatus, implement, machine, contrivance, implant, or other similar or related article, including a component part or accessory which is intended for use in the diagnosis of disease or other conditions or in the cure, mitigation, treatment or prevention of disease in humans or in other animals (particularly in mammals); is intended to affect the structure or any function of the body of a human or other animal; or is recognized in the official National Formulary or the United States Pharmacopoeia or any supplement to them. In particular, "medical device" includes a manufactured product which is used to cope with disease (particularly human disease), such as by preventing, diagnosing, treating, alleviating and or monitoring disease; care for injuries (particularly human injuries), such as by diagnosing, treating, alleviating, monitoring or compensating for injuries; meet anatomical needs (particularly human anatomical needs), such as by investigating, replacing, modifying or supporting anatomical structures; maintain physiological functions (particularly human physiological functions), such as by investigating, replacing, modifying or supporting physiological functions; support or sustain life (particularly human life); and control conception (particularly human conception).

The term "implantable medical device" refers to a medical device that is partly or totally inserted into the subject's body (such as the body of a human or other mammal) or a natural orifice thereof and is expected to stay there for an extended period of time (for example about thirty days or more) or is used to replace an epithelial surface or the surface of the eye and is expected to stay in use for an extended period of time (for example about thirty days or more).

Typically, surgical or medical procedures are used to insert or apply implantable medical devices, and surgical or medical procedures are used to remove them.

Tissue can be damaged, and inflammation can occur, during the use of medical devices during medical procedures, and from the implantation of medical devices. Under physiological conditions, tissue injury is sensed by stem cells which may be distant and migrate to the site of damage, which then undergo alternate stem cell differentiation and promote structural and functional tissue repair. The accumulation of redox-active transition coppers, particularly copper (II), in tissues is accompanied by a suppression of the normal tissue regeneration effected by stem cells. Elevated tissue levels of redox-active transitional coppers, particularly copper (II), suppress these normal biological behaviors of such undifferentiated cells.

A reduction in extracellular copper or extracellular copper concentrations is advantageous in that it will lead to either a reduction in copper mediated (direct or indirect) tissue damage and/or to improvement in tissue repair and/or inflammation. It is also believed to aid in the restoration of normal tissue stem cell responses. Reduction in extracellular copper or extracellular copper concentrations is particularly advantageous for the treatment of diseases, disorders, and/or conditions of the heart. See e.g. U.S. Patent Nos. 6,951,890, 6,348,465, and 6,610,693.

An increase in intracellular copper is advantageous in that it will lead to a decrease in oxidative damage and an increase in cytochrome c oxidase activity which, for example, can reduce inappropriate apoptosis and tissue degradation. See e.g. U.S. Patent Application 60/735,688, filed November 9, 2005.

A reduction in transition copper or transition copper concentrations, for example, copper (II) or copper (II) concentrations, will be advantageous in the treatment of tissue damage which results from medical procedures or implants and which may further be exacerbated by mechanisms that may be affected by or are dependent on excess copper or copper concentrations. A reduction in transition copper or transition copper concentrations, for example, a reduction in copper (II) or copper (II) concentrations, will also be advantageous in preventing the adverse effects that result from devices used in medical procedures, particularly adverse effects on tissue which comes into contact with medical devices, including, for example, medical implants.

For example, a reduction in copper or copper concentrations, particularly a reduction in copper (II) or copper (II) concentrations, will be advantageous in preventing or providing a reduction in and/or reversal of copper associated damage. This may include not only the prevention or lessening or reversal of tissue damage and inflammation, but also improved tissue repair by, for example, restoration of normal tissue stem cell responses. It will be appreciated that a reduction in redox-active transition coppers may be effected systemically, so that the localized administration of a copper antagonist, by for example, use or insertion of a medical device comprising a copper antagonist as described herein, may lead to lessened tissue damage and/or inflammation and/or enhanced tissue repair not only at or near the site of use or insertion, but elsewhere in the body.

Mammals that may be treated using the described and claimed devices and methods include, for example, a human being having, or at risk for developing, for example, tissue damage and/or organ dysfunction, for example cardiovascular tissue damage and/or dysfunction, and/or inflammation.

Mammals, including human beings, that may be treated using the described and claimed devices and methods include those that have or are at risk for developing undesired * copper levels, for example, copper (II) levels, including copper levels that can cause or lead to tissue damage, or lowered tissue repair response, including but not limited to vessel damage and repair, and inflammation. Treatment includes, for example, therapies to ameliorate and/or reverse, in whole or in part, damage resulting from diseases, disorders or conditions that are characterized in any part by copper-involved or mediated damage of tissue and/or vasculature, whether directly or indirectly, and/or to copper-involved or mediated impairment of normal tissue stem cell responses, whether directly or indirectly, and/or to inflammation. The invention has application in treating, inter alia, for example, heart failure, macrovascular disease or damage, microvascular disease or damage, and/or toxic {e.g., hypertensive) tissue and/or organ disease or damage (including such ailments as may, for example, be characterized by heart failure, cardiomyopathy, myocardial infarction, and related arterial and organ diseases), and inflammation, by administration of a copper antagonist(s).

Suitable copper antagonists include compounds of Formula I and II as described herein.

Compounds suitable as copper antagonists include cyclic and acyclic compounds according to Formula I:

FORMULA I wherein Xi, X₂, X₃ and X₄ are independently selected from the group consisting of N, S and O; R_1; R_2, R_3, R_4, R₅ and R₆ are independently selected from the group consisting of H, C₁ to C₁₀ straight chain or branched alkyl, C3 to ClO cycloalkyl, Cl to C6 alkyl C3 to ClO cycloalkyl, anyl, anyl substituted with 1 to 5 substituents, heteroaryl, fused aryl, Cl to C6 alkyl aryl, Cl to C6 alkyl aryl substituted with 1 to 5 substituents, Cl to C 5 alkyl heteroaryl, Cl to C6 alkyl fused aryl, -CH₂COOH, -CH₂SO₃H, -CH₂PO(OH)₂, and -CH₂P(CH₃)O(OH); nl, n2 and n3 are independently 2 or 3 and each of R₇₎ R_8, R_9, Rio_, Rn and R_J2 is independently selected and is selected from the group consisting of H, Cl to ClO straight chain or branched alkyl, C3 to ClO cycloalkyl, Cl to C6 alkyl, C3 to ClO cycloalkyl, aryl, aryl substituted with 1 to 5 substituents, heteroaryl fused aryl,

Cl to C6 alkyl aryl, Cl to C6 alkyl aryl substituted with 1 to 5 substituents, Cl to C 5 alkyl heteroaryl, Cl to C6 fused aryl, provided that when Xi is S or O, then R₂ is absent; when X₂ is S or O, then R₃ is absent, when X₃ is S or O, then R₄ is absent and when X₄ is S or O, then R₅ is absent.

Optionally, one or more of Ri_, R_2, R_3, R_4, R₅ and R₆ may be functionalized for attachment to groups which include, but are not limited to peptides, proteins, polyethylene glycols (PEGs) and other suitable chemical entities in order to modify the overall pharmacokinetics, deliverability and/or half lives of the constructs. Examples of such functionalization include, but are not limited to, Cl to ClO alkyl-CO-peptide, Cl to ClO alkyl-CO-protein, Cl to ClO alkyl-CO-PEG, Cl to ClO alkyl-NH-peptide, Cl to ClO alkyl-NH-protein, Cl to ClO alkyl-NH-CO-PEG, Cl to ClO alkyl-S-peptide, and Cl to ClO alkyl-S-protein. In addition, optionally are one or more of R₇, R₈, R₉, Rio, Rn and Ri₂ may be functionalized for attachment to groups which include, but are not limited to, peptides, proteins, polyethylene glycols (PEGs) and other suitable chemical entities in order to modify the overall pharmacokinetics, deliverability and/or half lives of the constructs. Examples of such functionalization include, but are not limited to, Cl to ClO alkyl-CO-peptide, Cl to ClO alkyl-CO-protein, Cl to ClO alkyl-CO-PEG, Cl to ClO alkyl-NH-peptide, Cl to ClO alkyl-NH-protein, Cl to ClO alkyl-NH-CO-PEG, Cl to ClO alkyl-S-peptide and Cl to ClO alkyl-S-protein.

One group of suitable compounds of Formula I include those wherein Ri₅R_2, R_3, R_4, R₅ and R₆ are independently selected from H, Cl to C6 alkyl, -CH₂COOH₅ -CH₂SO₃H, -CH₂PO(OH)₂ and -CH₂P(CH₃)O(OH); and each R_7, R_8, R_9, R_10, Rn and Ri₂ is independently selected from H and Cl to C6 alkyl. In one aspect, suitable compounds include those wherein at least one of Ri and R₂ and at least one of R₅ and R₆ is H or Cl to C6 alkyl. According to this aspect, suitably R₃ and R₄ are selected from H or Cl to C6 alkyl; more particularly, Ri_, R_2, R_5, and R₆ are selected from H or Cl to C6 alkyl. One sub-group of suitable compounds include those wherein X₂ and X₃ are N and nl, n2 and n3 are 2, or nl and n3 are

2 and n2 is 3. In this sub-group, R_1, R_6, R_7, Rg_, R_9, R_1O, Rn_, and R_i2 are independently selected from H and Cl to C3 alkyl. According to another subgroup of suitable compounds, all of X_1, X_2, X_3, and X₄ are suitably N or, alternatively, one OfX₁ and X₄ is S and X₂ and X₃ are N or S.

Tetra-heteroatom acyclic compounds within Formula I are provided where

Xi, X₂, X₃, and X₄ are independently chosen from the atoms N, S or O, such that,

(a) for a four-nitrogen series, i.e., when X₁, X₂, X₃, and X₄ are N then: Ri,

R₂, R₃, R₄, R₅, and R₆ are independently chosen from H, CH₃, C2-C10 straight chain or branched alkyl, C3-C10 cycloalkyl, C1-C6 alkyl C3-C10 cycloalkyl, aryl, mono, di, tri, tetra and penta substituted aryl, heteroaryl, fused aryl, C1-C6 alkyl aryl, C1-C6 alkyl mono, di, tri, tetra and penta substituted aryl, C1-C5 alkyl heteroaryl, C1-C6 alkyl fused aryl, CH₂COOH, CH₂SO₃H, CH₂PO(OH)₂, CH₂P(CH₃)O(OH); nl, n2, and n3 are independently chosen to be 2 or 3; and, R₇, R₈, R₉, R₁₀, R₁₁, and R₁₂ are independently chosen from H, CH₃, C2-C10 straight chain or branched alkyl, C3-C10 cycloalkyl, C1-C6 alkyl C3-C10 cycloalkyl, aryl, mono, di, tri, tetra and penta substituted aryl, heteroaryl, fused aryl, C1-C6 alkyl aryl, C1-C6 alkyl mono, di, tri, tetra and penta substituted aryl, C1-C5 alkyl heteroaryl, C1-C6 alkyl fused aryl. In addition, one or several Of R₁, R₂, R₃, R₄, R₅, or R₆ may be functionalized for attachment, for example, to peptides, proteins, polyethylene glycols and other such chemical entities in order to modify the overall pharmacokinetics, deliverability and/or half lives of the constructs. Examples of such functionalization include but are not limited to Cl-ClO alkyl- CO-peptide, Cl-ClO alkyl-CO-protein, Cl-ClO alkyl-CO-PEG, Cl-ClO alkyl- NH-peptide, Cl-ClO alkyl-NH-protein, Cl-ClO alkyl-NH-CO-PEG, Cl-ClO alkyl-S-peptide, Cl-ClO alkyl-S-protein. Furthermore one or several of R₇, R₈, R₉, R₁₀, Rn, or R₁₂ may be functionalized for attachment, for example, to peptides!, proteins, polyethylene glycols and other such chemical entities in order to modify the overall pharmacokinetics, deliverability and/or half lives of the constructs. Examples of such functionalization include but are not limited to Cl- ClO alkyl-CO-peptide, Cl-ClO alkyl-CO-protein, Cl-ClO alkyl-CO-PEG, Cl- ClO alkyl-NH-peptide, Cl-ClO alkyl-NH-protein, Cl-ClO alkyl-NH-CO-PEG,

Cl-ClO alkyl-S-peptide, and Cl-ClO alkyl-S-protein.

(b) for a first three-nitrogen series, i.e., when X₁, X₂, X₃, are N and X₄ is S or O then: R₆ does not exist; R₁, R₂, R₃, R₄ and R₅ are independently chosen from H, CH₃, C2-C10 straight chain or branched alkyl, C3-C10 cycloalkyl, C1-C6 alkyl C3-C10 cycloalkyl, aryl, mono, di, tri, tetra and penta substituted aryl, heteroaryl, fused aryl, C1-C6 alkyl aryl, C1-C6 alkyl mono, di, tri, tetra and penta substituted aryl, C1-C5 alkyl heteroaryl, C1-C6 alkyl fused aryl, CH₂COOH, CH₂SO₃H, CH₂PO(OH)₂, CH₂P(CH₃)O(OH); nl, n2, and n3 are independently chosen to be 2 or 3; and, R₇, R₈, R₉, Ri₀, Rn₅ and Ri₂ are independently chosen from H, CH₃, C2-C10 straight chain or branched alkyl, C3- ClO cycloalkyl, C1-C6 alkyl C3-C10 cycloalkyl, aryl, mono, di, tri, tetra and penta substituted aryl, heteroaryl, fused aryl, C1-C6 alkyl aryl, C1-C6 alkyl mono, di, tri, tetra and penta substituted aryl, C1-C5 alkyl heteroaryl, C1-C6 alkyl fused aryl. In addition, one or several of R₁, R₂, R₃, R₄, or R₅ may be functionalized for attachment, for example, to peptides, proteins, polyethylene glycols and other such chemical entities in order to modify the overall pharmacokinetics, deliverability and/or half lives of the constructs. Examples of such functionalization include but are not limited to Cl-ClO alkyl-CO-peptide, Cl-ClO alkyl-CO-protein, Cl-ClO alkyl-CO-PEG, Cl-ClO alkyl-NH-peptide, Cl-ClO alkyl-NH-protein, Cl-ClO alkyl-NH-CO-PEG, Cl-ClO alkyl-S-peptide, Cl-ClO alkyl-S-protein. Furthermore one or several Of R₇, R₈, R₉, R₁₀, Rn, or R₁₂ may be functionalized for attachment, for example, to peptides, proteins, polyethylene glycols and other such chemical entities in order to modify the overall pharmacokinetics, deliverability and/or half lives of the constructs. Examples of such functionalization include but are not limited to Cl-ClO alkyl- CO-peptide, Cl-ClO alkyl-CO-protein, Cl-ClO alkyl-CO-PEG, Cl-ClO alkyl- NH-peptide, Cl-ClO alkyl-NH-protein, Cl-ClO alkyl-NH-CO-PEG, Cl-ClO alkyl-S-peptide, and Cl-ClO alkyl-S-protein. (c) for a second three-nitrogen series, i.e., when X₁, X₂, and X₄ are N and X₃ is O or S then: R₄ does not exist and Ri, R₂, R₃, R₅, and R₆ are independently chosen from H, CH₃, C2-C10 straight chain or branched alkyl, C3-C10 cycloalkyl, C1-C6 alkyl C3-C10 cycloalkyl, aryl, mono, di, tri, tetra and penta substituted aryl, heteroaryl, fused aryl, C1-C6 alkyl aryl, C1-C6 alkyl mono, di, tri, tetra and penta substituted aryl, C1-C5 alkyl heteroaryl, C1-C6 alkyl fused aryl, CH₂COOH, CH₂SO₃H, CH₂PO(OH)₂, CH₂P(CH₃)O(OH); nl, n2, and n3 are independently chosen to be 2 or 3; and, R₇, R₈, R₉, R₁O, Rn, and R₁₂ are independently chosen from H, CH₃, C2-C10 straight chain or branched alkyl, C3- ClO cycloalkyl, C1-C6 alkyl C3-C10 cycloalkyl, aryl, mono, di, tri, tetra and penta substituted aryl, heteroaryl, fused aryl, C1-C6 alkyl aryl, C1-C6 alkyl mono, di, tri, tetra and penta substituted aryl, C1-C5 alkyl heteroaryl, C1-C6 alkyl fused aryl. In addition, one or several of Ri, R₂, R₃, R₅, or R₆ may be functionalized for attachment, for example, to peptides, proteins, polyethylene glycols and other such chemical entities in order to modify the overall pharmacokinetics, deliverability and/or half lives of the constructs. Examples of such functionalization include but are not limited to Cl-ClO alkyl-CO-peptide, Cl-ClO alkyl-CO-protein, Cl-ClO alkyl-CO-PEG, Cl-ClO alkyl-NH-peptide, Cl-ClO alkyl-NH-protein, Cl-ClO alkyl-NH-CO-PEG, Cl-ClO alkyl-S-peptide, Cl-ClO alkyl-S-protein. Furthermore one or several Of R₇, R₈, R₉, Rio, Rn, or Ri₂ may be functionalized for attachment, for example, to peptides, proteins, polyethylene glycols and other such chemical entities in order to modify the overall pharmacokinetics, deliverability and/or half lives of the constructs. Examples of such functionalization include but are not limited to Cl-ClO alkyl- CO-peptide, Cl-ClO alkyl-CO-protein, Cl-ClO alkyl-CO-PEG, Cl-ClO alkyl- NH-peptide, Cl-ClO alkyl-NH-protein, Cl-ClO alkyl-NH-CO-PEG, Cl-ClO alkyl-S-peptide, and Cl-ClO alkyl-S-ρrotein.

(d) for a first two-nitrogen series, i.e., when X₂ and X₃ are N and Xi and X₄ are O or S then: Ri and R₆ do not exist; R₂, R₃, R₄, and R₅ are independently

' chosen from H, CH₃, C2-C10 straight chain or branched alkyl, C3-C10 cycloalkyl, C1-C6 alkyl C3-C10 cycloalkyl, aryl, mono, di, tri, tetra and penta substituted aryl, heteroaryl, fused aryl, C1-C6 alkyl aryl, C1-C6 alkyl mono, di, tri, tetra and penta substituted aryl, C1-C5 alkyl heteroaryl, C1-C6 alkyl fused aryl, CH₂COOH₅ CH₂SO₃H₅ CH₂PO(OH)₂, CH₂P(CH₃)O(OH); nl, n2₅ and n3 are independently chosen to be 2 or 3; and R₇, R₈, R₉, R₁₀, R₁₁, and R₁₂ are independently chosen from H, CH₃, C2-C10 straight chain or branched alkyl, C3- ClO cycloalkyl, C1-C6 alkyl C3-C10 cycloalkyl, aryl, mono, di, tri, tetra and penta substituted aryl, heteroaryl, fused aryl, C1-C6 alkyl aryl, C1-C6 alkyl mono, di, tri, tetra and penta substituted aryl, C1-C5 alkyl heteroaryl, C1-C6 alkyl fused aryl. In addition, one or several of R₂, R₃, R₄, or R₅ may be functionalized for attachment, for example, to peptides, proteins, polyethylene glycols and other such chemical entities in order to modify the overall pharmacokinetics, deliverability and/or half lives of the constructs. Examples of such functionalization include but are not limited to Cl-ClO alkyl-CO-peptide, Cl-ClO alkyl-CO-protein, Cl-ClO alkyl-CO-PEG, Cl-ClO alkyl-NH-peptide, Cl-ClO alkyl-NH-protein, Cl-ClO alkyl-NH-CO-PEG, Cl-ClO alkyl-S-peptide, Cl-ClO alkyl-S-protein. Furthermore one or several of R₇, R₈, R₉, R₁₀, R₁₁, or R₁₂ may be functionalized for attachment, for example, to peptides, proteins, polyethylene glycols and other such chemical entities in order to modify the overall pharmacokinetics, deliverability and/or half lives of the constructs. Examples of such functionalization include but are not limited to Cl-ClO alkyl- CO-peptide, Cl-ClO alkyl-CO-protein, Cl-ClO alkyl-CO-PEG, Cl-ClO alkyl- NH-peptide, Cl-ClO alkyl-NH-protein, Cl-ClO alkyl-NH-CO-PEG, Cl-ClO alkyl-S-peptide, and Cl-ClO alkyl-S-protein.

(e) for a second two-nitrogen series, i.e., when X₁ and X₃ are N and X₂ and X₄ are O or S then: R₃ and R₆ do not exist; R₁, R₂, R₄, and R₅ are independently chosen from H₅ CH₃, C2-C10 straight chain or branched alkyl, C3-C10 cycloalkyl, C1-C6 alkyl C3-C10 cycloalkyl, aryl, mono, di, tri, tetra and penta substituted aryl, heteroaryl, fused aryl, C1-C6 alkyl aryl, C1-C6 alkyl mono, di, tri, tetra and penta substituted aryl, C1-C5 alkyl heteroaryl, C1-C6 alkyl fused aryl, CH₂COOH₅ CH₂SO₃H₅ CH₂PO(OH)₂, CH₂P(CH₃)O(OH); nl, n2, and n3 are independently chosen to be 2 or 3; and R₇, R₈, R₉, R₁₀, R₁₁, and R₁₂ are independently chosen from H₅ CH₃, C2-C10 straight chain or branched alkyl, C3- ClO cycloalkyl, C1-C6 alkyl C3-C10 cycloalkyl, aryl, mono, di, tri, tetra and penta substituted aryl, heteroaryl, fused aryl, C1-C6 alkyl aryl, C1-C6 alkyl mono, di, tri, tetra and penta substituted aryl, C1-C5 alkyl heteroaryl, C1-C6 alkyl fused aryl. In addition, one or several of R₁, R₂, R₄, or R₅ may be functionalized for attachment, for example, to peptides, proteins, polyethylene glycols and other such chemical entities in order to modify the overall pharmacokinetics, deliverability and/or half lives of the constructs. Examples of such functionalization include but are not limited to Cl-ClO alkyl-CO-peptide, Cl-ClO alkyl-CO-protein, Cl-ClO alkyl-CO-PEG, Cl-ClO alkyl-NH-peptide, Cl-ClO alkyl-NH-protein, Cl-ClO alkyl-NH-CO-PEG, Cl-ClO alkyl-S-peptide, and Cl-ClO alkyl-S-protein. Furthermore one or several Of R₇, R₈, R₉, Rio, Rn, or Ri₂ may be functionalized for attachment, for example, to peptides, proteins, polyethylene glycols and other such chemical entities in order to modify the overall pharmacokinetics, deliverability and/or half lives of the constructs. Examples of such functionalization include but are not limited to Cl-ClO alkyl- CO-peptide, Cl-ClO alkyl-CO-protein, Cl-ClO alkyl-CO-PEG, Cl-ClO alkyl- NH-peptide, Cl-ClO alkyl-NH-protein, Cl-ClO alkyl-NH-CO-PEG, Cl-ClO alkyl-S-peptide, and Cl-ClO alkyl-S-protein.

(f) for a third two-nitrogen series, i.e., when Xi, and X₂ are N and X₃ and X₄ are O or S then: R₄ and R₆ do not exist; Ri, R₂, R₃, and R₅ are independently chosen from H, CH₃, C2-C10 straight chain or branched alkyl, C3-C10 cycloalkyl, C1-C6 alkyl C3-C10 cycloalkyl, aryl, mono, di, tri, tetra and penta substituted aryl, heteroaryl, fused aryl, C1-C6 alkyl aryl, C1-C6 alkyl mono, di, tri, tetra and penta substituted aryl, C1-C5 alkyl heteroaryl, C1-C6 alkyl fused aryl, CH₂COOH, CH₂SO₃H, CH₂PO(OH)₂, CH₂P(CH₃)O(OH); nl, n2, and n3 are independently chosen to be 2 or 3; and R₇, R₈, R₉, Rio, Rn, and Ri₂ are independently chosen from H, CH₃, C2-C10 straight chain or branched alkyl, C3- ClO cycloalkyl, C1-C6 alkyl C3-C10 cycloalkyl, aryl, mono, di, tri, tetra and penta substituted aryl, heteroaryl, fused aryl, C1-C6 alkyl aryl, C1-C6 alkyl mono, di, tri, tetra and penta substituted aryl, C1-C5 alkyl heteroaryl, C1-C6 alkyl fused aryl. In addition, one or several of Ri, R₂, R₃, or R₅ may be functionalized for attachment, for example, to peptides, proteins, polyethylene glycols and other such chemical entities in order to modify the overall pharmacokinetics, deliverability and/or half lives of the constructs. Examples of such functionalization include but are not limited to Cl-ClO alkyl-CO-peptide, Cl-ClO alkyl-CO-protein, Cl-ClO alkyl-CO-PEG, Cl-ClO alkyl-NH-peptide, Cl-ClO alkyl-NH-protein, Cl-ClO alkyl-NH-CO-PEG, Cl-ClO alkyl-S-peptide, and Cl-ClO alkyl-S-protein. Furthermore one or several of R₇, R₈, R₉, Rio, Rn, or Rn may be functionalized for attachment, for example, to peptides, proteins, polyethylene glycols and other such chemical entities in order to modify the overall pharmacokinetics, deliverability and/or half lives of the constructs. Examples of such functionalization include but are not limited to Cl-ClO alkyl- CO-peptide, Cl-ClO alkyl-CO-protein, Cl-ClO alkyl-CO-PEG, Cl-ClO alkyl- NH-peptide, Cl-ClO alkyl-NH-protein, Cl-ClO alkyl-NH-CO-PEG, Cl-ClO alkyl-S-peptide, and Cl-ClO alkyl-S-protein.

(g) for a fourth two-nitrogen series, i.e., when Xi and X₄ are N and X₂ and X₃ are O or S then: R₃ and R₄ do not exist; Ri, R₂, R₅ and R₆ are independently chosen from H, CH₃, C2-C10 straight chain or branched alkyl, C3-C10 cycloalkyl, C1-C6 alkyl C3-C10 cycloalkyl, aryl, mono, di, tri, tetra and penta substituted aryl, heteroaryl, fused aryl, C1-C6 alkyl aryl, C1-C6 alkyl mono, di, tri, tetra and penta substituted aryl, C1-C5 alkyl heteroaryl, C1-C6 alkyl fused aryl, CH₂COOH, CH₂SO₃H, CH₂PO(OH)₂, CH₂P(CH₃)O(OH); nl, n2, and n3 are independently chosen to be 2 or 3; and R₇, R₈, R₉, Ri₀, Rn, and R_J2 are independently chosen from H, CH₃, C2-C10 straight chain or branched alkyl, C3- ClO cycloalkyl, C1-C6 alkyl C3-C10 cycloalkyl, aryl, mono, di, tri, tetra and penta substituted aryl, heteroaryl, fused aryl, C1-C6 alkyl aryl, C1-C6 alkyl mono, di, tri, tetra and penta substituted aryl, C1-C5 alkyl heteroaryl, C1-C6 alkyl fused aryl. In addition, one or several of Ri, R₂, R₅, or R₆ may be functionalized for attachment, for example, to peptides, proteins, polyethylene glycols and other such chemical entities in order to modify the overall pharmacokinetics, deliverability and/or half lives of the constructs. Examples of such functionalization include but are not limited to Cl-ClO alkyl-CO-peptide, Cl-ClO alkyl-CO-protein, Cl-ClO alkyl-CO-PEG, Cl-ClO alkyl-NH-peptide, Cl-ClO alkyl-NH-protein, Cl-ClO alkyl-NH-CO-PEG, Cl-ClO alkyl-S-ρeρtide, and Cl-ClO alkyl-S-protein. Furthermore one or several of R₇, R₈, R₉, R₁O, Rn, or R₁₂ may be functionalized for attachment, for example, to peptides, proteins, polyethylene glycols and other such chemical entities in order to modify the overall pharmacokinetics, deliverability and/or half lives of the constructs. Examples of such functionalization include but are not limited to Cl-ClO alkyl- CO-peptide, Cl-ClO alkyl-CO-protein, Cl-ClO alkyl-CO-PEG, Cl-ClO alkyl- NH-peptide, Cl-ClO alkyl-NH-protein, Cl-ClO alkyl-NH-CO-PEG, Cl-ClO alkyl-S-peptide, and Cl-ClO alkyl-S-protein. Second, for a tetra-heteroatom series of cyclic analogues, one Of R₁ and R₂ and one of R₅ and R₆ are joined together to form the bridging group (CR₁₃R₁₄)_n4, and X₁, X₂, X₃, and X₄ are independently chosen from the atoms N, S or O such that,

(a) for a four-nitrogen series, i.e., when X₁, X₂, X₃, and X₄ are N then: R₂, R₃, R₄, and R₅ are independently chosen from H, CH₃, C2-C10 straight chain or branched alkyl, C3-C10 cycloalkyl, C1-C6 alkyl C3-C10 cycloalkyl, aryl, mono, di, tri, tetra and penta substituted aryl, heteroaryl, fused aryl, C1-C6 alkyl aryl, C1-C6 alkyl mono, di, tri, tetra and penta substituted aryl, C1-C5 alkyl heteroaryl, C1-C6 alkyl fused aryl, CH₂COOH, CH₂SO₃H, CH₂PO(OH)₂, CH₂P(CH₃)O(OH); nl, n2, n3, and n4 are independently chosen to be 2 or 3; and R₇, R₈, R₉, R₁₀, R₁₁, R₁₂, R₁₃ and R₁₄ are independently chosen from H, CH₃, C2- ClO straight chain or branched alkyl, C3-C10 cycloalkyl, C1-C6 alkyl C3-C10 cycloalkyl, aryl, mono, di, tri, tetra and penta substituted aryl, heteroaryl, fused aryl, C1-C6 alkyl aryl, C1-C6 alkyl mono, di, tri, tetra and penta substituted aryl, C1-C5 alkyl heteroaryl, C1-C6 alkyl fused aryl. In addition, one or several of R₂, R₃, R₄, or R₅ may be functionalized for attachment, for example, to peptides, proteins, polyethylene glycols and other such chemical entities in order to modify the overall pharmacokinetics, deliverability and/or half lives of the constructs. Examples of such functionalization include but are not limited to Cl-ClO alkyl- CO-peptide, Cl-ClO alkyl-CO-protein, Cl-ClO alkyl-CO-PEG, Cl-ClO alkyl- NH-ρeptide, Cl-ClO alkyl-NH-protein, Cl-ClO alkyl-NH-CO-PEG, Cl-ClO alkyl-S-peptide, Cl-ClO alkyl-S-protein. Furthermore one or several of R₇, Rg,

R₉, Rio, Rn, Ri₂, Ri₃ or Ri₄ may be functionalized for attachment, for example, to peptides, proteins, polyethylene glycols and other such chemical entities in order to modify the overall pharmacokinetics, deliverability and/or half lives of the constructs. Examples of such functionalization include but are not limited to Cl- ClO alkyl-CO-peptide, Cl-ClO alkyl-CO-protein, Cl-ClO alkyl-CO-PEG, Cl- ClO alkyl-NH-peptide, Cl-ClO alkyl-NH-protein, Cl-ClO alkyl-NH-CO-PEG, Cl-ClO alkyl-S-ρeptide, and Cl-ClO alkyl-S-protein.

(b) for a three-nitrogen series, i.e., when Xi, X₂, X₃, are N and X₄ is S or O then: R₅ does not exist; R₂, R₃, and R₄ are independently chosen from H, CH₃, C2-C10 straight chain or branched alkyl, C3-C10 cycloalkyl, C1-C6 alkyl C3- ClO cycloalkyl, aryl, mono, di, tri, tetra and penta substituted aryl, heteroaryl, fused aryl, C1-C6 alkyl aryl, C1-C6 alkyl mono, di, tri, tetra and penta substituted aryl, C1-C5 alkyl heteroaryl, C1-C6 alkyl fused aryl, CH₂COOH, CH₂SO₃H, CH₂PO(OH)₂, CH₂P(CH₃)O(OH); nl, n2, n3, and n4 are independently chosen to be 2 or 3; and R₇, R₈, R₉, Rio, Rn, R₁₂, R_B and Ri₄ are independently chosen from H, CH₃, C2-C10 straight chain or branched alkyl, C3- ClO cycloalkyl, C1-C6 alkyl C3-C10 cycloalkyl, aryl, mono, di, tri, tetra and penta substituted aryl, heteroaryl, fused aryl, C1-C6 alkyl aryl, C1-C6 alkyl mono, di, tri, tetra and penta substituted aryl, C1-C5 alkyl heteroaryl, C1-C6 alkyl fused aryl. In addition, one or several of R₂, R₃ or R₄ may be functionalized for attachment, for example, to peptides, proteins, polyethylene glycols and other such chemical entities in order to modify the overall pharmacokinetics, deliverability and/or half-lives of the constructs. Examples of such functionalization include but are not limited to Cl-ClO alkyl-CO-peptide, Cl-ClO alkyl-CO-protein, Cl-ClO alkyl-CO-PEG, Cl-ClO alkyl-NH-peptide, Cl-ClO alkyl-NH-protein, Cl-ClO alkyl-NH-CO-PEG, Cl-ClO alkyl-S-peptide, and Cl-ClO alkyl-S-protein. Furthermore one or several of R₇, Rs, R9, Rio, Rn, R₁₂, Ri₃ or Ri₄ may be functionalized for attachment, for example, to peptides, proteins, polyethylene glycols and other such chemical entities in order to modify the overall pharmacokinetics, deliverability and/or half lives of the constructs. Examples of such functionalization include but are not limited to Cl-ClO alkyl- CO-peptide, Cl-ClO alkyl-CO-protein, Cl-ClO alkyl-CO-PEG, Cl-ClO alkyl- NH-peptide, Cl-ClO alkyl-NH-protein, Cl-ClO alkyl-NH-CO-PEG, Cl-ClO alkyl-S-peptide, and Cl-ClO alkyl-S-protein. (c) for a first two-nitrogen series, i.e., when X₂ and X₃ are N and X₁ and X₄ are O or S then: R₂ and R₅ do not exist; R₃ and R₄ are independently chosen from H, CH₃, C2-C10 straight chain or branched alkyl, C3-C10 cycloalkyl, C1-C6 alkyl C3-C10 cycloalkyl, aryl, mono, di, tri, tetra and penta substituted aryl, heteroaryl, fused aryl, C1-C6 alkyl aryl, C1-C6 alkyl mono, di, tri, tetra and penta substituted aryl, C1-C5 alkyl heteroaryl, C1-C6 alkyl fused aryl, CH₂COOH, CH₂SO₃H, CH₂PO(OH)₂, CH₂P(CH₃)O(OH); nl, n2, n3, and n4 are independently chosen to be 2 or 3; and R₇, Rg, R₉, R₁₀, R_11? Ri_2> R-₁3 and R₁₄ are independently chosen from H, CH₃, C2-C10 straight chain or branched alkyl, C3- ClO cycloalkyl, C1-C6 alkyl C3-C10 cycloalkyl, aryl, mono, di, tri, tetra and penta substituted aryl, heteroaryl, fused aryl, C1-C6 alkyl aryl, C1-C6 alkyl mono, di, tri, tetra and penta substituted aryl, C1-C5 alkyl heteroaryl, C1-C6 alkyl fused aryl. In addition, one or both of R₃, or R₄ may be functionalized for attachment, for example, to peptides, proteins, polyethylene glycols and other such chemical entities in order to modify the overall pharmacokinetics, deliverability and/or half-lives of the constructs. Examples of such functionalization include but are not limited to Cl-ClO alkyl-CO-peptide, Cl- ClO alkyl-CO-protein, Cl-ClO alkyl-CO-PEG, Cl-ClO alkyl-NH-peptide, Cl- ClO alkyl-NH-protein, Cl-ClO alkyl-NH-CO-PEG, Cl-ClO alkyl-S-peptide, and Cl-ClO alkyl-S-protein. Furthermore one or several of R₇, R₈, R₉, R₁₀, R₁₁, R₁₂, Ri₃ or R₁₄ may be functionalized for attachment, for example, to peptides, proteins, polyethylene glycols and other such chemical entities in order to modify the overall pharmacokinetics, deliverability and/or half lives of the constructs. Examples of such functionalization include but are not limited to Cl-ClO alkyl- CO-peptide, Cl-ClO alkyl-CO-protein, Cl-ClO alkyl-CO-PEG, Cl-ClO alkyl- NH-peptide, Cl-ClO alkyl-NH-protein, Cl-ClO alkyl-NH-CO-PEG, Cl-ClO alkyl-S-peptide, and Cl-ClO alkyl-S-protein. (d) for a second two-nitrogen series, i.e., when X₁ and X₃ are N and X₂ and

X₄ are O or S then: R₃ and R₅ do not exist; R₂ and R₄ are independently chosen from H, CH₃, C2-C10 straight chain or branched alkyl, C3-C10 cycloalkyl, Cl- C6 alkyl C3-C10 cycloalkyl, aryl, mono, di, tri, tetra and penta substituted aryl, heteroaryl, fused aryl, C1-C6 alkyl aryl, C1-C6 alkyl mono, di, tri, tetra and penta substituted aryl, C1-C5 alkyl heteroaryl, C1-C6 alkyl fused aryl, CH₂COOH, CH₂SO₃H, CH₂PO(OH)₂, CH₂P(CH₃)O(OH); nl, n2, n3, and n4 are independently chosen to be 2 or 3; and R₇, R₈, R₉, R₁₀, Rn, R₁₂, R₁₃ and R₁₄ are independently chosen from H, CH₃, C2-C10 straight chain or branched alkyl, C3- ClO cycloalkyl, C1-C6 alkyl C3-C10 cycloalkyl, aryl, mono, di, tri, tetra and penta substituted aryl, heteroaryl, fused aryl, C1-C6 alkyl aryl, C1-C6 alkyl mono, di, tri, tetra and penta substituted aryl, C1-C5 alkyl heteroaryl, C1-C6 alkyl fused aryl. In addition, one or both of R₂, or R₄ may be functionalized for attachment, for example, to peptides, proteins, polyethylene glycols and other such chemical entities in order to modify the overall pharmacokinetics, deliverability and/or half-lives of the constructs. Examples of such functionalization include but are not limited to Cl-ClO alkyl-CO-peptide, Cl- ClO alkyl-CO-protein, Cl-ClO alkyl-CO-PEG, Cl-ClO alkyl-NH-peptide, Cl- ClO alkyl-NH-protein, Cl-ClO alkyl-NH-CO-PEG, Cl-ClO alkyl-S-peptide, and Cl-ClO alkyl-S-protein. Furthermore one or several Of R₇, R₈, R₉, R₁₀, R₁₁, Ri₂, Ri₃ or R₁₄ may be functionalized for attachment, for example, to peptides, proteins, polyethylene glycols and other such chemical entities in order to modify the overall pharmacokinetics, deliverability and/or half lives of the constructs. Examples of such functionalization include but are not limited to Cl-ClO alkyl- CO-peptide, Cl-ClO alkyl-CO-protein, Cl-ClO alkyl-CO-PEG, Cl-ClO alkyl- NH-peptide, Cl-ClO alkyl-NH-protein, Cl-ClO alkyl-NH-CO-PEG, Cl-ClO alkyl-S-peptide, and Cl-ClO alkyl-S-protein.

(e) for a one-nitrogen series, i.e., when Xi is N and X₂, X₃ and X₄ are O or S then: R₃, R₄ and R₅ do not exist; R₂ is independently chosen from H, CH₃, C2- ClO straight chain or branched alkyl, C3-C10 cycloalkyl, C1-C6 alkyl C3-C10 cycloalkyl, aryl, mono, di, tri, tetra and penta substituted aryl, heteroaryl, fused aryl, C1-C6 alkyl aryl, C1-C6 alkyl mono, di, tri, tetra and penta substituted aryl,

C1-C5 alkyl heteroaryl, C1-C6 alkyl fused aryl, CH₂COOH, CH₂SO₃H, CH₂PO(OH)₂, CH₂P(CH₃)O(OH); nl, n2, n3, and n4 are independently chosen to be 2 or 3; and R₇, Rg, R₉, R₁O, Rn, Ri_2> R13 and R₁₄ are independently chosen from H, CH₃, C2-C10 straight chain or branched alkyl, C3-C10 cycloalkyl, Cl- C6 alkyl C3-C10 cycloalkyl, aryl, mono, di, tri, tetra and penta substituted aryl, heteroaryl, fused aryl, C1-C6 alkyl aryl, C1-C6 alkyl mono, di, tri, tetra and penta substituted aryl, C1-C5 alkyl heteroaryl, C1-C6 alkyl fused aryl. In addition, R₂ may be functionalized for attachment, for example, to peptides, proteins, polyethylene glycols and other such chemical entities in order to modify the overall pharmacokinetics, deliverability and/or half lives of the constructs. Examples of such functionalization include but are not limited to Cl-ClO alkyl- CO-peptide, Cl-ClO alkyl-CO-protein, Cl-ClO alkyl-CO-PEG, Cl-ClO alkyl- NH-peptide, Cl-ClO alkyl-NH-protein, Cl-ClO alkyl-NH-CO-PEG, Cl-ClO alkyl-S-peptide, and Cl-ClO alkyl-S-protein. Furthermore one or several of R₇, R₈, R₉, R₁₀, R₁₁, R₁₂, R₁₃ or R₁₄ may be functionalized for attachment, for example, to peptides, proteins, polyethylene glycols and other such chemical entities in order to modify the overall pharmacokinetics, deliverability and/or half lives of the constructs. Examples of such functionalization include but are not limited to Cl-ClO alkyl-CO-peptide, Cl-ClO alkyl-CO-protein, Cl-ClO alkyl- CO-PEG, Cl-ClO alkyl-NH-peptide, Cl-ClO alkyl-NH-protein, Cl-ClO alkyl- NH-CO-PEG, Cl-ClO alkyl-S-peptide, and Cl-ClO alkyl-S-protein.

Suitable copper antagonist compounds of Formula I include, for example:

SH-CH₂-CH₂-NH-CH₂-CH₂-NH-CH₂-CH₂-NH₂, SH-CH₂-CH₂-S-CH₂-CH₂-NH-CH₂-CH₂-NH₂, NH₂-CH₂-CH₂-NH-CH₂-CH₂-S-CH₂-CH₂-SH, NH₂-CH₂-CH₂-S-CH₂-CH₂-S-CH₂-CH₂-SH,

SH-CH₂-CH₂-S-CH₂-CH₂-S-CH₂-CH₂-SH, NH₂-CH₂-CH₂-NH-CH₂-CH₂-CH₂-NH-CH₂-CH₂-NH₂, SH-CH₂-CH₂-NH-CH₂-CH₂-CH₂-NH-CH₂-CH₂-NH₂, SH-CH₂-CH₂-S-CH₂-CH₂-CH₂-NH-CH₂-CH₂-NH₂, NH₂-CH₂-CH₂-NH-CH₂-CH₂-CH₂-S-CH₂-CH₂-SH,

NH₂-CH₂-CH₂-S-CH₂-CH₂-CH₂-S-CH₂-CH₂-SH, and SH-CH₂-CH₂-S-CH₂-CH₂-CH₂-S-CH₂-CH₂-SH.

Suitable compounds of Formula I include, for example, one or more of triethylenetetramine, salts of triethylenetetramine, prodrugs of triethylenetetramine and salts of such prodrugs; analogs of triethylenetetramine and salts and prodrugs of such analogs; and/or active metabolites of triethylenetetramine and salts and prodrugs of such metabolites, including but not limited to N-acetyl triethylenetetramine and salts and prodrugs of N-acetyl triethylenetetramine. Triethylenetetramine is a strongly basic moiety with multiple nitrogens that can be converted into a large number of suitable associated acid addition salts using an acid, for example, by reaction of triethylenetetramine and of the acid, for example, stoichiometrically equivalent amounts, in a solvent, for example, an inert solvent such as, for example, ethanol or water and subsequent evaporation if the dosage form is best formulated from a dry salt. Possible acids for this reaction are in particular those that yield physiologically acceptable salts. Nitrogen-containing copper chelator(s) or binding compound(s), for example, triethylenetetramine active agents such as, for example, triethylenetetramine, that can be delivered as a salt(s) (such as acid addition salts, e.g., triethylenetetramine dihydrochloride or triethylenetetramine disuccinate or other acceptable hydrochloride or succinate salts), act as copper-chelating or binding agents, which aids the elimination of copper from the body by forming a stable soluble complex that is readily excreted by the kidney. Thus, inorganic acids can be used, e.g., sulfuric acid, nitric acid, hydrohalic acids such as hydrochloric acid or hydrobromic acid, phosphoric acids such as orthophosphoric acid, and sulfamic acid. This is not an exhaustive list. Other organic acids can be used to prepare suitable salt forms, in particular aliphatic, alicyclic, araliphatic, aromatic or heterocyclic mono-or polybasic carboxylic, sulfonic or sulfuric acids, (e.g., formic acid, acetic acid, propionic acid, pivalic acid, diethylacetic acid, malonic acid, succinic acid, pimelic acid, fumaric acid, maleic acid, lactic acid, tartaric acid, malic acid, citric acid, gluconic acid, ascorbic acid, nicotinic acid, isonicotinic acid, methane-or ethanesulfonic acid, ethanedisulfonic acid, 2- hydroxyethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, naphthalenemono-and-disulfonic acids, and laurylsulfuric acid). Those in the art will be able to prepare other suitable salt forms. Nitrogen-containing copper chelator(s) or binding compound(s), for example, triethylenetetramine active agents such as, for example, triethylenetetramine, can also be in the form of quarternary ammonium salts in which the nitrogen atom carries a suitable organic group such as an alkyl, alkenyl, alkynyl or aralkyl moiety. In one embodiment, such nitrogen-containing copper chelator(s) are in the form of a compound or buffered in solution and/or suspension nearer to a neutral pH, lower than the pH 14 of a solution of triethylenetetramine itself.

Other triethylenetetramine active agents include derivative triethylenetetramine active agents, for example, triethylenetetramine in combination with picolinic acid (2-pyridinecarboxylic acid). These derivatives include, for example, triethylenetetramine picolinate and salts of triethylenetetramine picolinate, for example, triethylenetetramine picolinate HCl. These also include, for example, triethylenetetramine di-picolinate and salts of triethylenetetramine di-picolinate, for example, triethylenetetramine di-picolinate HCl. Picolinic acid moieties may be attached to triethylenetetramine, for example, one or more of the CH₂ moieties, using chemical techniques known in the art. Those in the art will be able to prepare other suitable derivatives, for example, triethylenetetramine-PEG derivatives, which may be useful for particular dosage forms including oral dosage forms having increased bioavailability.

Additional compounds suitable as copper antagonists include cyclic and 5 acyclic compounds according to Formula II:

FORMULA II wherein X₁, X₂ and X₃ are independently selected from the group consisting of N, S and O; R_1, R_2; R_3; R₅ and R₆ are independently selected from the group consisting of H, C₁ to C₁₀ straight chain or branched alkyl, C3 to ClO cycloalkyl,

15 Cl to C6 alkyl C3 to ClO cycloalkyl, anyl, anyl substituted with 1 to 5 substituents, heteroaryl, fused aryl, Cl to C6 alkyl aryl, Cl to C6 alkyl aryl substituted with 1 to 5 substituents, Cl to C5 alkyl heteroaryl, Cl to C6 alkyl fused aryl, -CH₂COOH, -CH₂SO₃H, -CH₂PO(OH)₂, and -CH₂P(CH₃)O(OH); nl and ii2 are independently 2 or 3 and each of R_7j Rg_, R₉ and Rio_, is independently

20 selected and is selected from the group consisting of H, Cl to ClO straight chain or branched alkyl, C3 to ClO cycloalkyl, Cl to C6 alkyl, C3 to ClO cycloalkyl, aryl, aryl substituted with 1 to 5 substituents, heteroaryl fused aryl, Cl to C6 alkyl aryl, Cl to C6 alkyl aryl substituted with 1 to 5 substituents, Cl to C5 alkyl heteroaryl, Cl to C6 fused aryl, provided that when Xi is S or O, then R₂ is

25 absent; when X₂ is S or O, then R₃ is absent, and when X₃ is S or O, then R₅ is absent.

Optionally, one or more of R_1, R_2; R_3j R₅ and R₆ may be functionalized for attachment to groups which include, but are not limited to peptides, proteins, polyethylene glycols (PEGs) and other suitable chemical entities in order to

30 modify the overall pharmacokinetics, deliverability and/or half lives of the constructs. Examples of such functionalization include, but are not limited to, Cl to ClO alkyl-CO-peptide, Cl to ClO alkyl-CO-protein, Cl to ClO alkyl-CO-PEG, Cl to ClO alkyl-NH-peptide, Cl to ClO alkyl-NH-protein, Cl to ClO alkyl-NH-CO-PEG, Cl to ClO alkyl-S-peptide, and Cl to ClO alkyl-S-protein. In addition, optionally are one or more of R₇, R₈, R₉, and R₁O may be functionalized for attachment to groups which include, but are not limited to, peptides, proteins, polyethylene glycols (PEGs) and other suitable chemical entities in order to modify the overall pharmacokinetics, deliverability and/or half lives of the constructs. Examples of such functionalization include, but are not limited to, Cl to ClO alkyl-CO-peptide, Cl to ClO alkyl-CO-protein, Cl to ClO alkyl-CO-PEG, Cl to ClO alkyl-NH-peptide, Cl to ClO alkyl-NH-protein, Cl to ClO alkyl-NH-CO-PEG, Cl to ClO alkyl-S-peptide and Cl to ClO alkyl-S-protein.

One group of suitable compounds of Formula I include those wherein R₁, R₂, R₃, R₅ and R₆ are independently selected from H, Cl to C6 alkyl, -CH₂COOH, -CH₂SO₃H, -CH₂PO(OH)₂ and -CH₂P(CH₃)O(OH); and each R_7, R_8, R₉ and R₁₀ is independently selected from H and Cl to C6 alkyl. In one aspect, suitable compounds include those wherein at least one of R₁ and R₂ and at least one of R₅ and R₆ is H or Cl to C6 alkyl. According to this aspect, suitably R₃ is selected from H or Cl to C6 alkyl; more particularly, R₁, R₂, R_5, and R₆ are selected from H or Cl to C6 alkyl. One sub-group of suitable compounds include those wherein R_1, R_6, R_7, R_8, R₉ and R_10, are independently selected from H and Cl to C3 alkyl. According to another sub-group of suitable compounds, all OfX_1, X₂ and X₃ are suitably N or, alternatively, one OfX₁ and X₃ is S and X₂ are N or S.

Tri-heteroatom compounds within Formula II are provided where Xi, X₂, and X₃ are independently chosen from the atoms N, S or O such that,

(a) for a three-nitrogen series, when X₁, X₂, and X₃ are N then: R₁, R₂, R₃, R₅, and R₆ are independently chosen from H, CH₃, C2-C10 straight chain or branched alkyl, C3-C10 cycloalkyl, C1-C6 alkyl C3-C10 cycloalkyl, aryl, mono, di, tri, tetra and penta substituted aryl, heteroaryl, fused aryl, C1-C6 alkyl aryl, C1-C6 alkyl mono, di, tri, tetra and penta substituted aryl, C1-C5 alkyl heteroaryl, C1-C6 alkyl fused aryl, CH₂COOH, CH₂SO₃H, CH₂PO(OH)₂, CH₂P(CH₃)O(OH); nl, and n2 are independently chosen to be 2 or 3; and R₇, R₈, R₉, and R₁₀ are independently chosen from H, CH₃, C2-C10 straight chain or branched alkyl, C3-C10 cycloalkyl, C1-C6 alkyl C3-C10 cycloalkyl, aryl, mono, di, tri, tetra and penta substituted aryl, heteroaryl, fused aryl, C1-C6 alkyl aryl, C1-C6 alkyl mono, di, tri, tetra and penta substituted aryl, C1-C5 alkyl heteroaryl, C1-C6 alkyl fused aryl. In addition, one or several Of R₁, R₂, R₃, R5 or R₆ may be functionalized for attachment, for example, to peptides, proteins, polyethylene glycols and other such chemical entities in order to modify the overall pharmacokinetics, deliverability and/or half lives of the constructs. Examples of such functionalization include but are not limited to Cl-ClO alkyl- CO-peptide, Cl-ClO alkyl-CO-protein, Cl-ClO alkyl-CO-PEG, Cl-ClO alkyl- NH-peptide, Cl-ClO alkyl-NH-protein, Cl-ClO alkyl-NH-CO-PEG, Cl-ClO alkyl-S-peptide, and Cl-ClO alkyl-S-protein. Furthermore one or several of R₇, Rg, R₉, or Rio may be functionalized for attachment, for example, to peptides, proteins, polyethylene glycols and other such chemical entities in order to modify the overall pharmacokinetics, deliverability and/or half-lives of the constructs. Examples of such functionalization include but are not limited to Cl-ClO alkyl- CO-peptide, Cl-ClO alkyl-CO-protein, Cl-ClO alkyl-CO-PEG, Cl-ClO alkyl- NH-peptide, Cl-ClO alkyl-NH-protein, Cl-ClO alkyl-NH-CO-PEG, Cl-ClO alkyl-S-peptide, and Cl-ClO alkyl-S-protein.

(b) for a first two-nitrogen series, when Xi and X₂ are N and X₃ is S or O then: R₃ does not exist; Ri , R₂, R₅, and R₆ are independently chosen from H, CH₃, C2-C10 straight chain or branched alkyl, C3-C10 cycloalkyl, C1-C6 alkyl C3-C10 cycloalkyl, aryl, mono, di, tri, tetra and penta substituted aryl, heteroaryl, fused aryl, C1-C6 alkyl aryl, C1-C6 alkyl mono, di, tri, tetra and penta substituted aryl, C1-C5 alkyl heteroaryl, C1-C6 alkyl fused aryl, CH₂COOH, CH₂SO₃H, CH₂PO(OH)₂, CH₂P(CH₃)O(OH); nl, and n2 are independently chosen to be 2 or 3; and R₇, R₈, R₉, and Ri₀ are independently chosen from H, CH₃, C2-C10 straight chain or branched alkyl, C3-C10 cycloalkyl, C1-C6 alkyl C3-C10 cycloalkyl, aryl, mono, di, tri, tetra and penta substituted aryl, heteroaryl, fused aryl, C1-C6 alkyl aryl, C1-C6 alkyl mono, di, tri, tetra and penta substituted aryl, C1-C5 alkyl heteroaryl, C1-C6 alkyl fused aryl. In addition, one or several Of R₁, R₂, R₅ or R₆ may be functionalized for attachment, for example, to peptides, proteins, polyethylene glycols and other such chemical entities in order to modify the overall pharmacokinetics, deliverability and/or half lives of the constructs. Examples of such functionalization include but are not limited to Cl-ClO alkyl-CO-peptide, Cl-ClO alkyl-CO-protein, Cl-ClO alkyl-CO-PEG, Cl-ClO alkyl-NH-peptide, Cl-ClO alkyl-NH-protein, Cl-ClO alkyl-NH-CO- PEG, Cl-ClO alkyl-S-peptide, and Cl-ClO alkyl-S-protein. Furthermore one or several of R₇, Rg, R₉, or R_1O may be functionalized for attachment, for example, to peptides, proteins, polyethylene glycols and other such chemical entities in order to modify the overall pharmacokinetics, deliverability and/or half-lives of the constructs. Examples of such functionalization include but are not limited to Cl-ClO alkyl-CO-peptide, Cl-ClO alkyl-CO-protein, Cl-ClO alkyl-CO-PEG, Cl-ClO alkyl-NH-peptide, Cl-ClO alkyl-NH-protein, Cl-ClO alkyl-NH-CO- PEG, Cl-ClO alkyl-S-peptide, and Cl-ClO alkyl-S-protein.

(c) for a second, two-nitrogen series, when X₁ and X₂ are N and X₃ is O or S then: R₅ does not exist; R₁, R₂, R₃, and R₆ are independently chosen from H, CH₃, C2-C10 straight chain or branched alkyl, C3-C10 cycloalkyl, C1-C6 alkyl C3-C10 cycloalkyl, aryl, mono, di, tri, tetra and penta substituted aryl, heteroaryl, fused aryl, C1-C6 alkyl aryl, C1-C6 alkyl mono, di, tri, tetra and penta substituted aryl, C1-C5 alkyl heteroaryl, C1-C6 alkyl fused aryl, CH₂COOH, CH₂SO₃H, CH₂PO(OH)₂, CH₂P(CH₃)O(OH); nl and n2 are independently chosen to be 2 or 3; and R₇, R₈, R₉, and R₁₀ are independently chosen from H, CH₃, C2-C10 straight chain or branched alkyl, C3-C10 cycloalkyl, C1-C6 alkyl C3-C10 cycloalkyl, aryl, mono, di, tri, tetra and penta substituted aryl, heteroaryl, fused aryl, C1-C6 alkyl aryl, C1-C6 alkyl mono, di, tri, tetra and penta substituted aryl, C1-C5 alkyl heteroaryl, C1-C6 alkyl fused aryl. In addition, one or several OfR₁, R₂, R₅, or R₆ may be functionalized for attachment, for example, to peptides, proteins, polyethylene glycols and other such chemical entities in order to modify the overall pharmacokinetics, deliverability and/or half lives of the constructs. Examples of such functionalization include but are not limited to Cl-ClO alkyl-CO-peptide, Cl-ClO alkyl-CO-protein, Cl-ClO alkyl-CO-PEG, Cl-ClO alkyl-NH-peptide, Cl-ClO alkyl-NH-protein, Cl-ClO alkyl-NH-CO- PEG, Cl-ClO alkyl-S-peptide, and Cl-ClO alkyl-S-protein. Furthermore one or several of R₇, R₈, R₉, or R_1O may be functionalized for attachment, for example, to peptides, proteins, polyethylene glycols and other such chemical entities in order to modify the overall pharmacokinetics, deliverability and/or half-lives of the constructs. Examples of such functionalization include but are not limited to Cl-ClO alkyl-CO-peptide, Cl-ClO alkyl-CO-protein, Cl-ClO alkyl-CO-PEG, Cl-ClO alkyl-NH-peptide, Cl-ClO alkyl-NH-protein, Cl-ClO alkyl-NH-CO- PEG, Cl-ClO alkyl-S-peptide, and Cl-ClO alkyl-S-protein.

A series of tri-heteroatom cyclic analogues according to the above Formula II are provided in which Ri and R₆ are joined together to form the bridging group (CRπRi₂)_n3, and X₁, X₂ and X₃ are independently chosen from the atoms N, S or O such that:

(a) for a three-nitrogen series, when Xi, X₂, and X₃ are N then: R₂, R₃, and R₅ are independently chosen from H, CH₃, C2-C10 straight chain or branched alkyl, C3-C10 cycloalkyl, C1-C6 alkyl C3-C10 cycloalkyl, aryl, mono, di, tri, tetra and penta substituted aryl, heteroaryl, fused aryl, C1-C6 alkyl aryl, C1-C6 alkyl mono, di, tri, tetra and penta substituted aryl, C1-C5 alkyl heteroaryl, Cl- C6 alkyl fused aryl, CH₂COOH, CH₂SO₃H, CH₂PO(OH)₂, CH₂P(CH₃)O(OH); nl, n2, and n3 are independently chosen to be 2 or 3; and R₇, R₈, R₉, R_]0, R_n, and Ri₂ are independently chosen from H, CH₃, C2-C10 straight chain or branched alkyl, C3-C10 cycloalkyl, C1-C6 alkyl C3-C10 cycloalkyl, aryl, mono, di, tri, tetra and penta substituted aryl, heteroaryl, fused aryl, C1-C6 alkyl aryl, C1-C6 alkyl mono, di, tri, tetra and penta substituted aryl, C1-C5 alkyl heteroaryl, Cl- C6 alkyl fused aryl. In addition, one or several of R₂, R₃*, or R₅ may be functionalized for attachment, for example, to peptides, proteins, polyethylene glycols and other such chemical entities in order to modify the overall pharmacokinetics, deliverability and/or half lives of the constructs. Examples of such functionalization include but are not limited to Cl-ClO alkyl-CO-peptide,

Cl-ClO alkyl-CO-protein, Cl-ClO alkyl-CO-PEG, Cl-ClO alkyl-NH-peρtide₅ Cl-ClO alkyl-NH-protein, Cl-ClO alkyl-NH-CO-PEG, Cl-ClO alkyl-S-peptide, and Cl-ClO alkyl-S-protein. Furthermore one or several of R₇, R₈, R₉, Rio, Rn, or Ri₂ may be functionalized for attachment, for example, to peptides, proteins, polyethylene glycols and other such chemical entities in order to modify the overall pharmacokinetics, deliverability and/or half lives of the constructs. Examples of such functionalization include but are not limited to Cl-ClO alkyl- CO-peptide, Cl-ClO alkyl-CO-protein, Cl-ClO alkyl-CO-PEG, Cl-ClO alkyl- NH-peptide, Cl-ClO alkyl-NH-protein, Cl-ClO alkyl-NH-CO-PEG, Cl-ClO alkyl-S-peptide, and Cl-ClO alkyl-S-protein.

(b) for a two-nitrogen series, when Xi and X₂ are N and X₃ is S or O then: R₅ does not exist; R₂, and R₃ are independently chosen from H, CH₃, C2-C10 straight chain or branched alkyl, C3-C10 cycloalkyl, C1-C6 alkyl C3-C10 cycloalkyl, aryl, mono, di, tri, tetra and penta substituted aryl, heteroaryl, fused aryl, C1-C6 alkyl aryl, C1-C6 alkyl mono, di, tri, tetra and penta substituted aryl, C1-C5 alkyl heteroaryl, C1-C6 alkyl fused aryl, CH₂COOH, CH₂SO₃H, CH₂PO(OH)₂, CH₂P(CH₃)O(OH); nl, n2, and n3 are independently chosen to be 2 or 3; and R₇, R₈, R₉, Ri₀, Rn, and R_J2 are independently chosen from H, CH₃, C2-C10 straight chain or branched alkyl, C3-C10 cycloalkyl, C1-C6 alkyl C3- ClO cycloalkyl, aryl, mono, di, tri, tetra and penta substituted aryl, heteroaryl, fused aryl, C1-C6 alkyl aryl, C1-C6 alkyl mono, di, tri, tetra and penta substituted aryl, C1-C5 alkyl heteroaryl, C1-C6 alkyl fused aryl. In addition, one or both of R₂ or R₃ may be functionalized for attachment, for example, to peptides, proteins, polyethylene glycols and other such chemical entities in order to modify the overall pharmacokinetics, deliverability and/or half-lives of the constructs. Examples of such functionalization include but are not limited to Cl- ClO alkyl-CO-peptide, Cl-ClO alkyl-CO-protein, Cl-ClO alkyl-CO-PEG, Cl- ClO alkyl-NH-peptide, Cl-ClO alkyl-NH-protein, Cl-ClO alkyl-NH-CO-PEG, Cl-ClO alkyl-S-peptide, and Cl-ClO alkyl-S-protein. Furthermore one or several of R₇, R₈, R₉, R₁₀, R_n, or R₁₂ may be functionalized for attachment, for example, to peptides, proteins, polyethylene glycols and other such chemical entities in order to modify the overall pharmacokinetics, deliverability and/or half lives of the constructs. Examples of such functionalization include but are not limited to Cl-ClO alkyl-CO-peptide, Cl-ClO alkyl-CO-protein, Cl-ClO alkyl- CO-PEG, Cl-ClO alkyl-NH-peptide, Cl-ClO alkyl-NH-protein, Cl-ClO alkyl- NH-CO-PEG, Cl-ClO alkyl-S-peptide, and Cl-ClO alkyl-S-protein.

(c) for a one-nitrogen series, when X₁ is N and X₂ and X₃ are O or S then: R₃ and R₅ do not exist; R₂ is independently chosen, from H, CH₃, C2-C10 straight chain or branched alkyl, C3-C10 cycloalkyl, C1-C6 alkyl C3-C10 cycloalkyl, aryl, mono, di, tri, tetra and penta substituted aryl, heteroaryl, fused aryl, C1-C6 alkyl aryl, C1-C6 alkyl mono, di, tri, tetra and penta substituted aryl, C1-C5 alkyl heteroaryl, C1-C6 alkyl fused aryl, CH₂COOH, CH₂SO₃H, CH₂PO(OH)₂, CH₂P(CH₃)O(OH); nl, n2, and n3 are independently chosen to be 2 or 3; and R₇, R₈, R₉, R₁₀, Rn, and R₁₂ are independently chosen from H, CH₃, C2-C10 straight chain or branched alkyl, C3-C10 cycloalkyl, C1-C6 alkyl C3-C10 cycloalkyl, aryl, mono, di, tri, tetra and penta substituted aryl, heteroaryl, fused aryl, C1-C6 alkyl aryl, C1-C6 alkyl mono, di, tri, tetra and penta substituted aryl, C1-C5 alkyl heteroaryl, C1-C6 alkyl fused aryl. In addition, R₂ may be functionalized for attachment, for example, to peptides, proteins, polyethylene glycols and other such chemical entities in order to modify the overall pharmacokinetics, deliverability and/or half lives of the constructs. Examples of such functionalization include but are not limited to Cl-ClO alkyl-CO-peptide, Cl- ClO alkyl-CO-protein, Cl-ClO alkyl-CO-PEG, Cl-ClO alkyl-NH-peptide, Cl- ClO alkyl-NH-protein, Cl-ClO alkyl-NH-CO-PEG, Cl-ClO alkyl-S-peptide, and Cl-ClO alkyl-S-protein. Furthermore one or several of R₇, R₈, R₉, Ri₀, Rn, or R₁₂ may be functionalized for attachment, for example, to peptides, proteins, polyethylene glycols and other such chemical entities in order to modify the overall pharmacokinetics, deliverability and/or half lives of the constructs. Examples of such functionalization include but are not limited to Cl-ClO alkyl- CO-peptide, Cl-ClO alkyl-CO-protein, Cl-ClO alkyl-CO-PEG, Cl-ClO alkyl- NH-peptide, Cl-ClO alkyl-NH-protein, Cl-ClO alkyl-NH-CO-PEG, Cl-ClO alkyl-S-peptide, and Cl-ClO alkyl-S-protein.

Copper antagonists useful in the invention also include copper chelators that have been pre-complexed with a non-copper metal ion prior to administration for therapy. Metal ions used for pre-complexing have a lower association constant for the copper antagonist than that of copper. For example, a metal ion for pre- complexing a copper antagonist that chelates Cu²⁺ is one that has a lower binding affinity for the copper antagonist than Cu²⁺. Preferred metal ions for precomplexing include calcium (e.g., Ca²⁺), magnesium (e.g., Mg²⁺), chromium (e.g., Cr and Cr ), manganese (e.g., Mn ), zinc (e.g., Zn ), selenium (e.g., Se⁴⁺), and iron (e.g., Fe²⁺ and Fe³⁺). Most preferred metal ions for precomplexing are calcium, zinc, and iron. Other metals include, for example, cobalt (e.g., Co²⁺), nickel (e.g., Ni²⁺), silver (e.g., Ag¹⁺), and bismuth (e.g., Bi³⁺). Metals are chosen with regard, for example, to their relative binding to the copper antagonist, and relative to toxicity and the dose of the copper antagonist to be administered.

Also encompassed are metal complexes comprising copper antagonists and non^Acopper metals (that have lower binding affinities than copper for the copper antagonist) and one or more additional ligands than typically found in complexes of that metal. These additional ligands may serve to block sites of entry into the complex for water, oxygen, hydroxide, or other species that may undesirably complex with the metal ion and can cause degradation of the copper antagonist. For example, copper complexes of triethylenetetramine have been found to form pentacoordinate complexes with a tetracoordinated triethylenetetramine and a chloride ligand when crystallized from a salt solution rather than a tetracoordinate Cu²⁺ triethylenetetramine complex. In this regard, 219 mg of triethylenetetramine ^• 2 HCl were dissolved in 50 ml, and 170 mg of CuCl₂ • 2H2O were dissolved in 25 ml ethanol (95%). > After addition of the CuCl₂ solution to the triethylenetetramine solution, the color changed from light to dark blue and white crystals precipitated. The crystals were dissolved by addition of a solution of 80 mg NaOH in 15 ml H2O. After the solvent was evaporated, the residue was dissolved in ethanol, and two equivalents of ammonium- hexafluorophosphate were added. Blue crystals could be obtained after reduction of the solvent. Crystals were found that were suitable for x-ray structure determination. X-ray crystallography revealed a [Cu(triethylenetetramine)Cl] complex. Other coordinated complexes may be formed from or between copper antagonists, for example, copper chelators (such as Cu2+ chelators, spermidine, spermine, tetracyclam, etc.), particularly those subject to degradative pathways such as those noted above, by providing additional complexing agents (such as anions in solution, for example, I^", Br^", F^", (SO₄)^2", (CO₃)^2", BF^4", NO^3", ethylene, pyridine, etc.) in solutions of such complexes. This may be particularly desirable for complexes with more accessible metal ions, such as planar complexes or complexes having four or fewer coordinating agents, where one or more additional complexing agents could provide additional shielding to the metal from undesirable ligands that might otherwise access the metal and displace a desired complexing agent.

The compounds for use according to the present invention, including triethylenetetramine active agents, may be made using any of a variety of chemical synthesis, isolation, and purification methods known in the art. Exemplary synthetic routes are described below. General synthetic chemistry protocols are somewhat different for these classes of molecules due to their propensity to chelate with metallic cations, including copper. Glassware should be cleaned and silanized prior to use. Plasticware should be chosen specifically to have minimal presence of metal ions. Metal implements such as spatulas should be excluded from any chemistry protocol involving chelators. Water used should be purified by sequential carbon filtering, ion exchange and reverse osmosis to the highest level of purity possible, not by distillation. All organic solvents used should be rigorously purified to exclude any possible traces of metal ion contamination.

Care must also be take with purification of such derivatives due to their propensity to chelate with a variety of cations, including copper, which may be present in trace amounts in water, on the surface of glass or plastic vessels. Once again, glassware should be cleaned and silanized prior to use. Plasticware should be chosen specifically to have minimal presence of metal ions. Metal implements such as spatulas should be avoided, and water used should be purified by sequential carbon filtering, ion exchange and reverse osmosis to the highest level of purity possible, and not by distillation. All organic solvents used should be rigorously purified to exclude any possible traces of metal ion contamination. Ion exchange chromatography followed by lyophilization is typically the best way to obtain pure solid materials of these classes of molecules. Ion exchange resins should be washed clean of any possible metal contamination.

Many of the synthetic routes allow for control of the particular R groups introduced. For synthetic methods incorporating amino acids, synthetic amino acids can be used to incorporate a variety of substituent R groups. The dichloroethane synthetic schemes also allow for the incorporation of a wide variety of R groups by using dichlorinated ethane derivatives. It will be appreciated that many of these synthetic schemes can lead to isomeric forms of the compounds; such isomers can be separated using techniques known in the art.

Documents describing aspects of these synthetic schemes include the following: (1) A W von Hoffman, Berichte 23, 3711 (1890); (2) The Polymerization Of Ethylenimine, Giffin D. Jones, Arne Langsjoen, Sister Mary Marguerite Christine Neumann, Jack Zomlefer, J. Org. Chem., 1944; 9(2); 125- 147; (3) The peptide way to macrocyclic bifunctional chelating agents: synthesis of 2-(p-nitrobenzyl)- 1 ,4,7, 10-tetraazacyclododecane-N,N' ,N",N' "-tetraacetic acid and study of its yttrium(III) complex, Min K. Moi et al, J. Am. Chem. Soc, 1988; 770(18); 6266-6267; (4) Synthesis of a kinetically stable ⁹⁰Y labelled macrocycle-antibody conjugate, Jonathan P L Cox, et al., J. Chem. Soc. Chem. Comm., 797 (1989); (5) Specific and stable labeling of antibodies with technetium-99m with a diamide dithiolate chelating agent, Fritzberg AR, Abrams PG, Beaumier PL, Kasina S, Morgan AC, Rao TN, Reno JM, Sanderson JA, Srinivasan A, Wilbur DS, et al., Proc. Natl. Acad. Sc.i U. S. A. 85(11):4025-4029 (1988 Jun); (6) Towards tumour imaging with ¹¹¹In labelled macrocycle-antibody conjugates, Andrew S Craig et at, J. Chem. Soc. Chem. Comm., 794 (1989); (7)

Synthesis of C- and N-functionalised derivatives of NOTA, DOTA, and DTPA: bifunctional complexing agents for the derivitisation of antibodies, Jonathan P L Cox et at,, J. Chem. Soc. Perkin. I, 2567 (1990); (8) Macrocyclic chelators as anticancer agents in radioimmunotherapy, N R A Beeley and P R J Ansell, Current Opinions in Therapeutic Patents, 2:1539-1553 (1992); and (9) Synthesis of new macrocyclic amino-phosphinic acid complexing agents and their C- and P- functionalised derivatives for protein linkage, Christopher J Broan et at, Synthesis, 63 (1992). Acyclic and cyclic compounds of the invention and exemplary synthetic methods and existing syntheses from the art include the following: For tetra-heteroatom acyclic examples of Formula I:

X₁, X₂, X₃, and X₄ are independently chosen from the atoms N, S or O such that:

4N series: when X₁, X₂, X₃, and X₄ are N then:

R₁, R₂, R₃, R₄, R₅, and R₆ are independently chosen from H, CH₃, C2- ClO straight chain or branched alkyl, C3-C10 cycloalkyl, C1-C6 alkyl C3-C10 cycloalkyl, aryl, mono, di, tri, tetra and penta substituted aryl, heteroaryl, fused aryl, C1-C6 alkyl aryl, C1-C6 alkyl mono, di, tri, tetra and penta substituted aryl, C1-C5 alkyl heteroaryl, C1-C6 alkyl fused aryl, CH₂COOH, CH₂SO₃H, CH₂PO(OH)₂, CH₂P(CH₃)O(OH); nl, n2, and n3 are independently chosen to be 2 or 3, and each repeat of any of nl, n2, and n3 may be the same as or different than any other repeat; and R₇, R₈, R₉, R₁₀, Rn, and R_ϊ2 are independently chosen from H, CH₃,

C2-C10 straight chain or branched alkyl, C3-C10 cycloalkyl, C1-C6 alkyl C3- ClO cycloalkyl, aryl, mono, di, tri, tetra and penta substituted aryl, heteroaryl, fused aryl, C1-C6 alkyl aryl, C1-C6 alkyl mono, di, tri, tetra and penta substituted aryl, C1-C5 alkyl heteroaryl, C1-C6 alkyl fused aryl.

In addition, one or several of Ri, R₂, R₃, R₄, R₅, or R₆ may be functionalized for attachment, for example, to peptides, proteins, polyethylene glycols and other such chemical entities in order to modify the overall pharmaco-kinetics, deliverability and/or half lives of the constructs. Examples of such functionalization include but are not limited to Cl-ClO alkyl-CO-peptide, Cl- ClO alkyl-CO-protein, Cl-ClO alkyl-CO-PEG, Cl-ClO alkyl-NH-peptide, Cl- ClO alkyl-NH-protein, Cl-ClO alkyl-NH-CO-PEG, Cl-ClO alkyl- S-peptide, and Cl-ClO alkyl-S-protein.

Furthermore one or several of R₇, Rg, R₉, Rio, Rn, or Ri₂ may be functionalized for attachment, for example, to peptides, proteins, polyethylene glycols and other such chemical entities in order to modify the overall pharmaco- kinetics, deliverability and/or half lives of the constructs. Examples of such functionalization include but are not limited to Cl-ClO alkyl-CO-peptide, Cl- ClO alkyl-CO-protein, Cl-ClO alkyl-CO-PEG, Cl-ClO alkyl-NH-peptide, Cl- ClO alkyl-NH-protein, Cl-ClO alkyl-NH-CO-PEG, Cl-ClO alkyl-S-peptide, and Cl-ClO alkyl-S-ρrotein.

Also provided are embodiments wherein one, two, three or four of R₁ through Ri₂ are other than hydrogen.

In some embodiments, the compounds of Formula I or II are selective for a particular oxidation state of copper. For example, the compounds may be selected so that they preferentially bind oxidized copper, or copper (II). Copper selectivity can be assayed using methods known in the art. Competition assays can be done using isotopes of copper (I) and copper (II) to determine the ability of the compounds to selectively bind one form of copper. In some embodiments, the compounds of Formula I or II may be chosen to avoid excessive lipophilicity, for example by avoiding large or numerous alkyl substituents. Excessive lipophilicity can cause the compounds to bind to and/or pass through cellular membranes, thereby decreasing the amount of compound available for chelating copper, particularly for extracellular copper, which may be predominantly in the oxidized form of copper (II). Synthesis of examples of the open chain 4N series of Formula I

Triethylenetetramine itself has been synthesized by reaction of 2 equivalents of ethylene diamine with 1,2-dichloro ethane to give triethylenetetramine directly (1). Modification of this procedure by using starting materials with appropriate R_a and R_b groups (where R_a, R_b = R₇, R₈ or R₁₁, R₁₂) would lead to symmetrically substituted open chain 4N examples as shown below:

The judicious use of protecting group chemistry such as the widely used BOC (t-butyloxycarbonyl) group allows the chemistry to be directed specifically towards the substitution pattern shown. Other approaches such as via the chemistry of ethyleneimine (2) may also lead to a subset of the tetra-aza series. In order to obtain the un-symmetrically substituted derivatives a variant of some chemistry described by Meares et al (3) should be used. Standard peptide synthesis using the Rink resin along with FMOC protected natural and un-natural amino acids which can be conveniently cleaved at the penultimate step of the synthesis generates a tri-peptide C-terminal amide. This is reduced using Diborane in THF to give tlie open chain tetra-aza compounds as shown below:

The incorporation of Ri, R₂, R₅ and R₆ can be accomplished with this chemistry by standard procedures.

Resin

The reverse Rink approach, shown above, also leads to this class of tetra- aza derivatives and may be useful in cases where peptide coupling of a sterically hindered amino acid requires multiple coupling attempts in order to achieve success in the initial Rink approach.

The oxalamide approach, shown above, also can lead to successful syntheses of this class of compounds, although the central substituents are always going to be hydrogen or its isotopes with this kind of chemistry. This particular variant makes use of the trichloroethyl ester group to protect one of the carbolxylic acid functions of oxalic acid but other protecting groups are also envisaged. Reaction of an amino acid amide derived from a natural or unnatural amino acid with a differentially protected oxalyl mono chloride gives the mono- oxalamide shown which can be reacted under standard peptide coupling condition to give the un-symmetrical bis-oxalamide which can then be reduced with diborane to give the desired tetra-aza derivative.

3NX series 1: when X₁, X₂, X₃, are N and X₄ is S or O then:

R₆ does not exist

R₁, R₂, R₃, R₄ and R₅ are independently chosen from H, CH₃, C2-C10 straight chain or branched alkyl, C3-C10 cycloalkyl, C1-C6 alkyl C3-C10 cycloalkyl, aryl, mono, di, tri, tetra and penta substituted aryl, heteroaryl, fused aryl, C1-C6 alkyl aryl, C1-C6 alkyl mono, di, tri, tetra and penta substituted aryl, C1-C5 alkyl heteroaryl, C1-C6 alkyl fused aryl, CH₂COOH, CH₂SO₃H, CH₂PO(OH)₂, CH₂P(CH₃)O(OH); nl, n2, and n3 are independently chosen to be 2 or 3, and each repeat of any of nl, n2, and n3 may be the same as or different than any other repeat; and R₇, R₈, R₉, R_1O, Rn, and R_ϊ2 are independently chosen from H, CH₃,

In addition, one or several OfR₁, R₂, R₃, R₄, or R₅ may be functionalized for attachment, for example, to peptides, proteins, polyethylene glycols and other such chemical entities in order to modify the overall pharmacokinetics, deliverability and/or half lives of the constructs. Examples of such functionalization include but are not limited to Cl-ClO alkyl-CO-peptide, Cl- ClO alkyl-CO-protein, Cl-ClO alkyl-CO-PEG, Cl-ClO alkyl-NH-peptide, Cl- ClO alkyl-NH-protein, Cl-ClO alkyl-NH-CO-PEG, Cl-ClO alkyl- S-peptide, Cl- ClO alkyl-S-protein.

Furthermore one or several of R₇, R₈, R₉, R₁₀, Rn, or Rj₂ may be functionalized for attachment, for example, to peptides, proteins, polyethylene glycols and other such chemical entities in order to modify the overall pharmacokinetics, deliverability and/or half lives of the constructs. Examples of such functionalization include but are not limited to Cl-ClO alkyl-CO-peptide, Cl- ClO alkyl-CO-protein, Cl-ClO alkyl-CO-PEG, Cl-ClO alkyl-NH-peptide, Cl- ClO alkyl-NH-protein, Cl-ClO alkyl-NH-CO-PEG, Cl-ClO alkyl- S-peptide, Cl- ClO alkyl-S-protein.

Synthesis of examples of the open chain 3NX series 1 of Formula I: Variations of the syntheses used for the 4N series provide examples of the 3N series 1 class of compounds. The chemistry described by Meares et al (3) can be modified to give examples of the 3NX series of compounds.

Standard peptide synthesis according to the so-called reverse Rink approach as shown above using FMOC protected natural and un-natural amino acids which can be conveniently cleaved at the penultimate step of the synthesis generates a modified tri-peptide C-terminal amide. The cases where X₄ is O are incorporated by the use of an alpha-substituted carboxylic acid in the last coupling step. This is reduced using Diborane in THF to give the open chain tetra-aza compounds.

The incorporation of R₁, R₂, R₅ and R₆ can be accomplished with this chemistry by standard procedures.

Resin →

For the cases where X₄ = S a similar approach using standard peptide synthesis according to the so-called reverse Rink approach as shown above can be used. Coupling with FMOC protected natural and un-natural amino acids, which can be conveniently cleaved at the penultimate step of the synthesis, generates a modified tri-peptide C-terminal amide. The incorporation of X₄ = S is achieved by the use of an alpha-substituted carboxylic acid in the last coupling step. This is reduced using Diborane in THF to give the open chain tetra-aza compounds.

The oxalamide approach, shown above, can also lead to successful syntheses of this class of compounds, although the central substituents are always going to be hydrogen or its isotopes with this kind of chemistry. This particular variant makes use of the trichloroethyl ester group to protect one of the carbolxylic acid functions of oxalic acid but other protecting groups are also envisaged. Reaction of an amino acid amide derived from a natural or unnatural amino acid with a differentially protected oxalyl mono chloride gives the mono- oxalamide shown which can be reacted under standard peptide coupling conditions with an ethanolamine or ethanethiolamine derivative to give the un- symmetrical bis-oxalamide which can then be reduced with diborane as shown to give the desired tri-aza derivative.

3NX series 2: when Xi, X₂, and X₄ are N and X₃ is O or S then: R₄ does not exist, and

Ri, R₂, R₃, R₅, and R₆ are independently chosen from H, CH₃, C2-C10 straight chain or branched alkyl, C3-C10 cycloalkyl, C1-C6 alkyl C3-C10 cycloalkyl, aryl, mono, di, tri, tetra and penta substituted aryl, heteroaryl, fused aryl, C1-C6 alkyl aryl, C1-C6 alkyl mono, di, tri, tetra and penta substituted aryl, C1-C5 alkyl heteroaryl, C1-C6 alkyl fused aryl, CH₂COOH, CH₂SO₃H, CH₂PO(OH)₂, CH₂P(CH₃)O(OH); nl, n2, and n3 are independently chosen to be 2 or 3, and each repeat of any of nl, n2, and n3 may be the same as or different than any other repeat; and

R₇, Rg, R₉, Rio, Rn, and Ri₂ are independently chosen from H, CH₃,

In addition, one or several of Ri, R₂, R₃, R₅, or R₆ may be functionalized for attachment, for example, to peptides, proteins, polyethylene glycols and other such chemical entities in order to modify the overall pharmaco-kinetics, deliverability and/or half lives of the constructs. Examples of such functionalization include but are not limited to Cl-ClO alkyl-CO-peptide, Cl-

ClO alkyl-CO-protein, Cl-ClO alkyl-CO-PEG, Cl-ClO alkyl-NH-peptide, Cl-

ClO alkyl-NH-protein, Cl-ClO alkyl-NH-CO-PEG, Cl-ClO alkyl-S-peptide, Cl- ClO alkyl-S-protein.

Furthermore one or several of R₇, R₈, R₉, Rio, Rn, or R_i2 may be functionalized for attachment, for example, to peptides, proteins, polyethylene glycols and other such chemical entities in order to modify the overall pharmacokinetics, deliverability and/or half lives of the constructs. Examples of such functionalization include but are not limited to Cl-ClO alkyl-CO-peptide, Cl- ClO alkyl-CO-protein, Cl-ClO alkyl-CO-PEG, Cl-ClO alkyl-NH-peptide, Cl- ClO alkyl-NH-protein, Cl-ClO alkyl-NH-CO-PEG, Cl-ClO alkyl-S-peptide, and ^! Cl-ClO alkyl-S-ρrotein.

Synthesis of examples of the open chain 3NX series 2 of Formula I: A different approach can be used for the synthesis of the 3N series 2 class of compounds. The key component is the incorporation in the synthesis of an appropriately substituted and protected ethanolamine or ethanethiolamine derivative, which is readily available from both natural and un-natural amino acids, as shown below.

X₃ = O or S

The BOC protected ethanolamine or ethanethiolamine is reacted with an appropriate benzyl protected alpha chloroacid. After hydrogenation to deprotect the ester function, standard peptide coupling with a natural or unnatural amino acid amide followed by deprotection and reduction with diborane in THF gives the open chain tri-aza compounds. If hydrogenation is not compatible with other functionality in the molecule then alternative combinations of protecting groups can be used such as trichloroethyloxy carbonyl and t-butyl.

The incorporation of R₁, R₂, R₅ and R₆ can be accomplished with this chemistry by standard procedures. 2N2X series 1: when X₂ and X₃ are N and X₁ and X₄ are O or S then: Ri and R₆ do not exist; R₂, R₃, R₄, and R₅ are independently chosen from H, CH₃, C2-C10 straight chain or branched alkyl, C3-C10 cycloalkyl, C1-C6 alkyl C3-C10 cycloalkyl, aryl, mono, di, tri, tetra and penta substituted aryl, heteroaryl, fused aryl, C1-C6 alkyl aryl, C1-C6 alkyl mono, di, tri, tetra and penta substituted aryl, C1-C5 alkyl heteroaryl, C1-C6 alkyl fused aryl, CH₂COOH, CH₂SO₃H, CH₂PO(OH)₂, CH₂P(CH₃)O(OH); nl, n2, and n3 are independently chosen to be 2 or 3, and each repeat of any of nl, n2, and n3 may be the same as or different than any other repeat; and R₇, R₈, R₉, R_1O, Rn, and R₁₂ are independently chosen from H, CH₃,

C2-C10 straight chain or branched alkyl, C3-C10 cycloalkyl, C1-C6 alkyl C3- ClO cycloalkyl, aryl, mono, di, tri, tetra and penta substituted aryl, heteroaryl, fused aryl, C1-C6 alkyl aryl, C1-C6 alkyl mono, di, tri, tetra and penta substituted aryl, C1-C5 alkyl heteroaryl, C1-C6 alkyl fused aryl. In addition, one or several of R₂, R₃, R₄, or R₅ may be functionalized for attachment, for example, to peptides, proteins, polyethylene glycols and other such chemical entities in order to modify the overall pharmaco-kinetics, deliverability and/or half lives of the constructs. Examples of such functionalization include but are not limited to Cl-ClO alkyl-CO-peptide, Cl- ClO alkyl-CO-protein, Cl-ClO alkyl-CO-PEG, Cl-ClO alkyl-NH-peptide, Cl- ClO alkyl-NH-protein, Cl-ClO alkyl-NH-CO-PEG, Cl-ClO alkyl- S-peptide, and Cl-ClO alkyl-S-protein.

Furthermore one or several of R₇, R₈, R₉, R₁₀, Rn, or R₁₂ may be functionalized for attachment, for example, to peptides, proteins, polyethylene glycols and other such chemical entities in order to modify the overall pharmaco-kinetics, deliverability and/or half lives of the constructs. Examples of such functionalization include but are not limited to Cl-ClO alkyl-CO-peptide, Cl- ClO alkyl-CO-protein, Cl-ClO alkyl-CO-PEG, Cl-ClO alkyl-NH-peptide, Cl- ClO alkyl-NH-protein, Cl-ClO alkyl-NH-CO-PEG, Cl-ClO alkyl-S-peptide, Cl- ClO alkyl-S-protein. Synthesis of examples of the open chain 2N2X series 1 of Formula

I:

The oxalamide approach, shown above, can lead to successful syntheses of this class of compounds. This particular variant makes use of the trichloroethyl ester group to protect one of the carbolxylic acid functions of oxalic acid but other protecting groups are also envisaged. Reaction of an aminoalcohol or aminothiol derivative readily available from a natural or unnatural amino acid with a differentially protected oxalyl mono chloride gives the mono-oxalamide shown which can be reacted under standard peptide coupling condition to give the un-symmetrical bis-oxalamide which can then be reduced with diborane to give the desired tetra-aza derivative.

X₁ = O or S

X₄ = O or S

A variant of the dichloroethane approach, shown above, can also lead to successful syntheses of this class of compounds. Reaction of an aminoalcohol or aminothiol derivative readily available from a natural or unnatural amino acid with an O-protected 1-chloro, 2-hydroxy ethane derivative followed by deprotection and substitution with chloride gives the mono-chloro compound shown which can be further reacted with an appropriate aminoalcohol or aminothiol derivative readily available from a natural or unnatural amino acid to give the un-symmetrical desired product. 2N2X series 2: when X₁ and X₃ are N and X₂ and X₄ are O or S then: R₃ and R₆ do not exist;

R₁, R₂, R₄, and R₅ are independently chosen from H, CH₃, C2-C10 straight chain or branched alkyl, C3-C10 cycloalkyl, C1-C6 alkyl C3-C10 cycloalkyl, aryl, mono, di, tri, tetra and penta substituted aryl, heteroaryl, fused aryl, C1-C6 alkyl aryl, C1-C6 alkyl mono, di, tri, tetra and penta substituted aryl, C1-C5 alkyl heteroaryl, C1-C6 alkyl fused aryl, CH₂COOH, CH₂SO₃H, CH₂PO(OH) ₂, CH₂P(CH₃)O(OH); nl, n2, and n3 are independently chosen to be 2 or 3, and each repeat of any of nl, n2, and n3 may be the same as or different than any other repeat; and

R₇, Rg, R₉, R_1O, Rn, and R₁₂ are independently chosen from H, CH₃,

C2-C10 straight chain or branched alkyl, C3-C10 cycloalkyl, C1-C6 alkyl C3-

ClO cycloalkyl, aryl, mono, di, tri, tetra and penta substituted aryl, heteroaryl, fused aryl, C1-C6 alkyl aryl, C1-C6 alkyl mono, di, tri, tetra and penta substituted aryl, C1-C5 alkyl heteroaryl, C1-C6 alkyl fused aryl.

In addition, one or several of R₁, R₂, R₄, or R₅ may be functionalized for attachment, for example, to peptides, proteins, polyethylene glycols and other such chemical entities in order to modify the overall pharmaco-kinetics, deliverability and/or half lives of the constructs. Examples of such functionalization include but are not limited to Cl-ClO alkyl-CO-peptide, Cl- ClO alkyl-CO-protein, Cl-ClO alkyl-CO-PEG, Cl-ClO alkyl-NH-peptide, Cl- ClO alkyl-NH-protein, Cl-ClO alkyl-NH-CO-PEG, Cl-ClO alkyl-S-peptide, Cl- ClO alkyl-S-protein. Furthermore one or several of R₇, R₈, R₉, R₁₀, R₁₁, or R₁₂ may be functionalized for attachment, for example, to peptides, proteins, polyethylene glycols and other such chemical entities in order to modify the overall pharmacokinetics, deliverability and/or half lives of the constructs. Examples of such functionalization include but are not limited to Cl-ClO alkyl- CO-peptide, Cl- ClO alkyl-CO-protein, Cl-ClO alkyl-CO-PEG, Cl-ClO alkyl-NH-peptide, Cl- ClO alkyl-NH-protein, Cl-ClO alkyl-NH-CO-PEG, Cl-ClO alkyl-S-peptide, C 1 - ClO alkyl-S-protein.

Synthesis of the open chain 2N2X series 2 of Formula I:

S S

A variant of the dichloroethane approach, shown above, can lead to successful syntheses of this class of compounds. Reaction of an aminoalcohol or aminothiol derivative readily available from a natural or unnatural amino acid with an O-protected 1-chloro, 2-hydroxy ethane derivative followed by deprotection and substitution with chloride gives the mono-chloro compound shown which can be further reacted with an appropriately protected aminoalcohol or aminothiol derivative, readily available from a natural or unnatural amino acid, to give the un-symmetrical desired product after de- protection.

2N2X series 3: when Xi and X₂ are N and X₃ and X₄ are O or S then: R₄ and R₆ do not exist;

Ri, R₂, R₃, and R₅ are independently chosen from H, CH₃, C2-C10 straight chain or branched alkyl, C3-C10 cycloalkyl, C1-C6 alkyl C3-C10 cycloalkyl, aryl, mono, di, tri, tetra and penta substituted aryl, heteroaryl, fused aryl, C1-C6 alkyl aryl, C1-C6 alkyl mono, di, tri, tetra and penta substituted aryl, C1-C5 alkyl heteroaryl, C1-C6 alkyl fused aryl, CH₂COOH, CH₂SO₃H, CH₂PO(OH)₂, CH₂P(CH₃)O(OH); nl, n2, and n3 are independently chosen to be 2 or 3, and each repeat of any of nl, n2, and n3 may be the same as or different than any other repeat; and

R₇, R₈, R₉, R_1O, Rn, and R₁₂ are independently chosen from H, CH₃, C2-C10 straight chain or branched alkyl, C3-C10 cycloalkyl, C1-C6 alkyl C3- ClO cycloalkyl, aryl, mono, di, tri, tetra and penta substituted aryl, heteroaryl, fused aryl, C1-C6 alkyl aryl, C1-C6 alkyl mono, di, tri, tetra and penta substituted aryl, C1-C5 alkyl heteroaryl, C1-C6 alkyl fused aryl.

In addition, one or several of R₁, R₂, R₃, or R₅ may be functionalized for attachment, for example, to peptides, proteins, polyethylene glycols and other such chemical entities in order to modify the overall pharmaco-kinetics, deliverability and/or half lives of the constructs. Examples of such functionalization include but are not limited to Cl-ClO alkyl-CO-peptide, Cl- ClO alkyl-CO-protein, Cl-ClO alkyl-CO-PEG, Cl-ClO alkyl-NH-peptide, Cl- ClO alkyl-NH-protein, Cl-ClO alkyl-NH-CO-PEG, Cl-ClO alkyl-S-peptide, and Cl-ClO alkyl-S-protein.

Furthermore one or several of R₇, R₈, R₉, R₁₀, Rn, or R₁₂ may be functionalized for attachment, for example, to peptides, proteins, polyethylene glycols and other such chemical entities in order to modify the overall pharmacokinetics, deliverability and/or half lives of the constructs. Examples of such functionalization include but are not limited to Cl-ClO alkyl-CO-peptide, Cl- ClO alkyl-CO-protein, Cl-ClO alkyl-CO-PEG, Cl-ClO alkyl-NH-peptide, Cl- ClO alkyl-NH-protein, Cl-ClO alkyl-NH-CO-PEG, Cl-ClO alkyl-S-peptide, and Cl-ClO alkyl-S-protein. Synthesis of the open chain 2N2X series 3:

A variant of the dichloroethane approach, shown above, can lead to successful syntheses of this class of compounds. Reaction of a monoprotected ethylene diamine derivative, readily available from a natural or unnatural amino acid with an O-protected 1-chloro, 2-hydroxy ethane derivative followed by deprotection and substitution with chloride gives the mono-chloro compound shown which can be further reacted with an appropriately protected bis-alcohol or bis thiol derivative, readily available from a natural or unnatural amino acid, to give the un- symmetrical desired product after de-protection.

2N2X series 4: when Xi and X₄ are N and X₂ and X₃ are O or S then: R₃ and R₄ do not exist;

R₁, R₂, R₅ and R₆ are independently chosen from H, CH₃, C2-C10 straight chain or branched alkyl, C3-C10 cycloalkyl, C1-C6 alkyl C3-C10 cycloalkyl, aryl, mono, di, tri, tetra and penta substituted aryl, heteroaryl, fused aryl, C1-C6 alkyl aryl, C1-C6 alkyl mono, di, tri, tetra and penta substituted aryl, C1-C5 alkyl heteroaryl, C1-C6 alkyl fused aryl, CH₂COOH, CH₂SO₃H, CH₂PO(OH) ₂, CH₂P(CH₃)O(OH); nl, n2, and n3 are independently chosen to be 2 or 3, and each repeat of any of nl, n2, and n3 may be the same as or different than any other repeat; and R₇, R₈, R₉, R_1O, Ri_b and R₁₂ are independently chosen from H, CH₃,

In addition, one or several of R₁, R₂, R₅, or R₆ may be functionalized for attachment, for example, to peptides, proteins, polyethylene glycols and other such chemical entities in order to modify the overall pharmaco-kinetics, deliverability and/or half lives of the constructs. Examples of such functionalization include but are not limited to Cl-ClO alkyl-CO-peptide, Cl- ClO alkyl-CO-protein, Cl-ClO alkyl-CO-PEG, Cl-ClO alkyl-NH-peptide, Cl- ClO alkyl-NH-protein, Cl-ClO alkyl-NH-CO-PEG, Cl-ClO alkyl- S-peptide, and Cl-ClO alkyl-S-protein.

Furthermore one or several of R₇, R₈, R₉, Rio, Rn, or R₁₂ may be functionalized for attachment, for example, to peptides, proteins, polyethylene glycols and other such chemical entities in order to modify the overall pharmaco- kinetics, deliverability and/or half lives of the constructs. Examples of such functionalization include but are not limited to Cl-ClO alkyl-CO-peptide, Cl- ClO alkyl-CO-protein, Cl-ClO alkyl-CO-PEG, Cl-ClO alkyl-NH-peptide, Cl- ClO alkyl-NH-protein, Cl-ClO alkyl-NH-CO-PEG, Cl-ClO alkyl-S-peptide, and Cl-ClO alkyl-S-protein.

Synthesis of the open chain 2N2X series 4 of Formula I:

A variant of the dichloroethane approach, shown above, can lead to successful syntheses of this class of compounds. Reaction of a an appropriately protected bis-alcohol or bis thiol derivative, readily available from a natural or unnatural amino acid, with an O-protected 1-chloro, 2-hydroxy ethane derivative followed by deprotection and substitution with chloride gives the mono-chloro compound shown which can be further reacted with an appropriately protected bis-alcohol or bis thiol derivative, readily available from a natural or unnatural amino acid, to give the un-symmetrical desired product after de-protection.

For the Tetra-heteroatom cyclic series:

One of R₁ and R₂ (if Ri does not exist) and one of R₅ (if R₆ does not exist) and R₆ are joined together to form the bridging group (CR₁₃Ri₄)M;

Xi, X₂, X₃, and X₄ are independently chosen from the atoms N, S or O such that:

4N macrocyclic series: when X₁, X₂, X₃, and X₄ are N then:

R₂, R₃, R₄, and R₅ are independently chosen from H, CH₃, C2-C10 straight chain or branched alkyl, C3-C10 cycloalkyl, C1-C6 -alkyl C3-C10 cycloalkyl, aryl, mono, di, tri, tetra and penta substituted aryl, heteroaryl, fused aryl, C1-C6 alkyl aryl, C1-C6 alkyl mono, di, tri, tetra and penta substituted aryl, C1-C5 alkyl heteroaryl, C1-C6 alkyl fused aryl, CH₂COOH, CH₂SO₃H, CH₂PO(OH) ₂, CH₂P(CH₃)O(OH); nl, n2, n3, and n4 are independently chosen to be 2 or 3, and each repeat of any of nl, n2, n3 and n4 may be the same as or different than any other repeat; and

R₇, R₈, R₉, Rio, Rn, Ri₂, Ri₃ and Ri₄ are independently chosen from H,

CH₃, C2-C10 straight chain or branched alkyl, C3-C10 cycloalkyl, C1-C6 alkyl

C3-C10 cycloalkyl, aryl, mono, di, tri, tetra and penta substituted aryl, heteroaryl, fused aryl, C1-C6 alkyl aryl, C1-C6 alkyl mono, di, tri, tetra and penta substituted aryl, C1-C5 alkyl heteroaryl, C1-C6 alkyl fused aryl. In addition, one or several of R₂, R₃, R₄, or R₅ may be functionalized for attachment, for example, to peptides, proteins, polyethylene glycols and other such chemical entities in order to modify the overall pharmaco-kinetics, deliverability and/or half lives of the constructs. Examples of such functionalization include but are not limited to Cl-ClO alkyl-CO-peptide, Cl- ClO alkyl-CO-protein, Cl-ClO alkyl-CO-PEG, Cl-ClO alkyl-NH-peptide, Cl- ClO alkyl-NH-protein, Cl-ClO alkyl-NH-CO-PEG, Cl-ClO alkyl-S-peptide, and Cl-ClO alkyl-S-protein.

Furthermore one or several Of R₇, R₈, R₉, Rio, Rn, R12, R13 or Rw m^aY be functionalized for attachment, for example, to peptides, proteins, polyethylene glycols and other such chemical entities in order to modify the overall pharmaco- kinetics, deliverability and/or half lives of the constructs. Examples of such functionalization include but are not limited to Cl-ClO alkyl-CO-peptide, Cl- ClO alkyl-CO-protein, Cl-ClO alkyl-CO-PEG, Cl-ClO alkyl-NH-peptide, Cl- ClO alkyl-NH-protein, Cl-ClO alkyl-NH-CO-PEG, Cl-ClO alkyl-S-peptide, C 1 - ClO alkyl-S-ρrotein.

Synthesis of examples of the macrocyclic 4N series of Formula I:

Triethylenetetramine itself has been synthesized by reaction of 2 equivalents of ethylene diamine with 1,2-dichloro ethane to give triethylenetetramine directly

(1). Possible side products from this synthesis include the 12N4 macrocycle shown below, which could also be synthesized directly from

Triethylenetetramine by reaction with a further equivalent of 1,2-dichloro ethane under appropriately dilute concentrations to provide the 12N4 macrocycle shown. Modification of this procedure by using starting materials with appropriate R_a and R_b (where R_a> R_b correspond to R₇, R₈ or R₁₁, R₁₂) groups would lead to symmetrically substituted 12N4 macrocycle examples as shown below: 2 equivs

2 equ

The judicious use of protecting group chemistry such as the widely used

BOC (t-butyloxycarbonyl) group allows the chemistry to be directed specifically towards the substitution pattern shown. Other approaches such as via the chemistry of ethyleneimine (2) may also lead to a subset of the tetra-aza series. In order to obtain the un-symmetrically substituted derivatives a variant of some chemistry described by Meares et al (3) should be used. Standard peptide synthesis using the Merrifield approach or the SASRIN resin along with FMOC protected natural and un-natural amino acids which can be conveniently cleaved at a later step of the synthesis generates a fully protected tetra-peptide C-terminal SASRIN derivative. Cleavage of the N terminal FMOC protecting group followed by direct cyclization upon concomitant cleavage from the resin gives the macrocyclic tetrapeptide. This is reduced using Diborane in THF to give the 12N4 series of compounds as shown below: Resin

Resin

The reverse Merrifield/SASRIN approach, shown above, also leads to this class of tetra-aza derivatives and may be useful in cases where peptide coupling of a sterically hindered amino acid requires multiple coupling attempts in order to achieve success in the initial Merrifield approach.

Ri1.Ri2

The oxalamide approach, shown above, also can lead to successful syntheses of this class of compounds. This particular variant makes use of the trichloroethyl ester group to protect one of the carbolxylic acid functions of oxalic acid but other protecting groups are also envisaged. Reaction of an amino acid amide derived from a natural or unnatural amino acid with a differentially protected oxalyl mono chloride gives the mono-oxalamide shown which can be reacted under standard peptide coupling condition to give the un-symmetrical bis-oxalamide which can then be reduced with diborane to give the desired tetra- aza derivative. Further reaction with oxalic acid gives the cyclic derivative, which can then be reduced once again with diborane to give the 12N4 series of compounds.

3NX series: when X₁, X₂, X₃, are N and X₄ is S or O then:

R₅ does not exist;

R₂, R₃, and R₄ are independently chosen from H, CH₃, C2-C10 straight chain or branched alkyl, C3-C10 cycloalkyl, C1-C6 alkyl C3-C10 cycloalkyl, aryl, mono, di, tri, tetra and penta substituted aryl, heteroaryl, fused aryl, C1-C6 alkyl aryl, C1-C6 alkyl mono, di, tri, tetra and penta substituted aryl, C1-C5 alkyl heteroaryl, C1-C6 alkyl fused aryl, CH₂COOH, CH₂SO₃H, CH₂PO(OH) ₂,

CH₂P(CH₃)O(OH); nl, n2, n3, and n4 are independently chosen to be 2 or 3, and each repeat of any of nl, n2, n3 and n4 may be the same as or different than any other repeat; and

R₇, R₈, R₉, R_1O, R₁₁, Rn, Ri3 and R₁₄ are independently chosen from H,

CH₃, C2-C10 straight chain or branched alkyl, C3-C10 cycloalkyl, C1-C6 alkyl

C3-C10 cycloalkyl, aryl, mono, di, tri, tetra and penta substituted aryl, heteroaryl, fused aryl, C1-C6 alkyl aryl, C1-C6 alkyl mono, di, tri, tetra and penta substituted aryl, C 1 -C5 alkyl heteroaryl, C 1 -C6 alkyl fused aryl.

In addition, one or several of R₂, R₃ or R₄ may be functionalized for attachment, for example, to peptides, proteins, polyethylene glycols and other such chemical entities in order to modify the overall pharmaco-kinetics, deliverability and/or half lives of the constructs. Examples of such functionalization include but are not limited to Cl-ClO alkyl-CO-peptide, Cl- ClO alkyl-CO-protein, Cl-ClO alkyl-CO-PEG, Cl-ClO alkyl-NH-peptide, Cl- ClO alkyl-NH-protein, Cl-ClO alkyl-NH-CO-PEG, Cl-ClO alkyl- S-peptide, and Cl-ClO alkyl-S-protein.

Furthermore one or several of R₇, R₈, R₉, R₁₀, R₁₁, R₁₂, R₁₃ or R₁₄ may be functionalized for attachment, for example, to peptides, proteins, polyethylene glycols and other such chemical entities in order to modify the overall pharmacokinetics, deliverability and/or half lives of the constructs. Examples of such functionalization include but are not limited to Cl-ClO alkyl-CO-peptide, Cl- ClO alkyl-CO-protein, Cl-ClO alkyl-CO-PEG, Cl-ClO alkyl-NH-peptide, Cl- ClO alkyl-NH-protein, Cl-ClO alkyl-NH-CO-PEG, Cl-ClO alkyl- S-peptide, Cl- ClO alkyl-S-ρrotein.

Synthesis of examples of the macrocyclic 3NX series of Formula I:

Triethylenetetramine itself has been synthesized by reaction of 2 equivalents of ethylene diamine with 1,2-dichloro ethane to give triethylenetetramine directly (1). Possible side products from this synthesis include the 12N4 macrocycle shown below, which could also be synthesized directly from Triethylenetetramine by reaction with a further equivalent of 1,2-dichloro ethane under appropriately dilute concentrations to provide the 12N4 macrocycle shown. Modification of this procedure by using starting materials with appropriate R groups leads to symmetrically substituted 12N4 macrocycle examples as shown below:

BOC

X₄ = O or S

The judicious use of protecting group chemistry such as the widely used

BOC (t-butyloxycarbonyl) group allows the chemistry to be directed specifically towards the substitution pattern shown. Other approaches such as via the chemistry of ethyleneimine (2) may also lead to a subset of the tri-aza X series. In order to obtain alternative un-symmetrically substituted derivatives a variant of some chemistry described by Meares et al (3) could be used. Standard peptide synthesis using the Merrifield approach or the SASRIN resin along with FMOC protected natural and un-natural amino acids which can be conveniently cleaved at a later step of the synthesis generates a tri-peptide C-terminal SASRTN derivative which can be further elaborated with an appropriate BOCO or BOCS compound the give the resin bound 3NX compound shown. Reduction with diborane followed by Tosylation would give the 3NX OTosyl linear compound, which, upon deprotection and cyclization would give the desired 3NX macrocycle as shown below:

^ReSin

BH₃ In THF Tosylation Resin *- »^►

BH₃ In THF Tosylation Resin

The reverse Merrifield/SASRTN approach, shown above, also leads to this class of tetra-aza derivatives and may be useful in cases where peptide coupling of a sterically hindered amino acid requires multiple coupling attempts in order to achieve success in the initial Merrifield approach. 2N2X series 1: when X₂ and X₃ are N and Xi and X₄ are O or S then: R₂ and R₅ do not exist

R₃ and R₄ are independently chosen from H, CH₃, C2-C10 straight chain or branched alkyl, C3-C10 cycloalkyl, C1-C6 alkyl C3-C10 cycloalkyl, aryl, mono, di, tri, tetra and penta substituted aryl, heteroaryl, fused aryl, C1-C6 alkyl aryl, C1-C6 alkyl mono, di, tri, tetra and penta substituted aryl, C1-C5 alkyl heteroaryl, C1-C6 alkyl fused aryl, CH₂COOH, CH₂SO₃H, CH₂PO(OH)₂, CH₂P(CH₃)O(OH); nl, n2, n3, and n4 are independently chosen to be 2 or 3, and each repeat of any of nl, n2, n3 and n4 may be the same as or different than any other repeat; and

R₇, R₈, R₉, R_1O, Rn, Ri_2> Rn and R₁₄ are independently chosen from H,

CH₃, C2-C10 straight chain or branched alkyl, C3-C10 cycloalkyl, C1-C6 alkyl

C3-C10 cycloalkyl, aryl, mono, di, tri, tetra and penta substituted aryl, heteroaryl, fused aryl, C1-C6 alkyl aryl, C1-C6 alkyl mono, di, tri, tetra and penta substituted aryl, C1-C5 alkyl heteroaryl, C1-C6 alkyl fused aryl

In addition, one or both of R₃, or R₄ may be functionalized for attachment, for example, to peptides, proteins, polyethylene glycols and other such chemical entities in order to modify the overall pharmaco-kinetics, deliverability and/or half lives of the constructs. Examples of such functionalization include but are not limited to Cl-ClO alkyl-CO-peptide, Cl-ClO alkyl-CO-protein, Cl-ClO alkyl-CO-PEG, Cl-ClO alkyl-NH-peptide, Cl-ClO alkyl-NH-protein, Cl-ClO alkyl-NH-CO-PEG, Cl-ClO alkyl- S-peptide, Cl-ClO alkyl- S -protein.

Furthermore one or several of R₇, R₈, R₉, R₁₀, Rn, R₁₂, Ri₃ or R₁₄ may be functionalized for attachment, for example, to peptides, proteins, polyethylene glycols and other such chemical entities in order to modify the overall pharmaco- kinetics, deliverability and/or half lives of the constructs. Examples of such functionalization include but are not limited to Cl-ClO alkyl-CO-peptide, Cl- ClO alkyl-CO-protein, Cl-ClO alkyl-CO-PEG, Cl-ClO alkyl-NH-peptide, Cl- ClO alkyl-NH-protein, Cl-ClO alkyl-NH-CO-PEG, Cl-ClO alkyl-S-peptide, C 1 - ClO alkyl-S-protein.

Synthesis of examples of the macrocyclic 2N2X series 1 of Formula I: '^■ .

The oxalamide approach, shown above, again can lead to successful syntheses of this class of compounds, although the central substituents are always going to be hydrogen or its isotopes with this kind of chemistry. This particular variant makes use of the trichloroethyl ester group to protect one of the carboxylic acid functions of oxalic acid but other protecting groups are also envisaged. Reaction of an aminoalcohol or aminothiol derivative readily available from a natural or unnatural amino acid with a differentially protected oxalyl mono chloride gives the mono-oxalamide shown which can be reacted under standard peptide coupling condition to give the un-symmetrical bis- oxalamide which can then be reduced with diborane to give the desired di-aza derivative. Deprotection followed by cyclization would give the 12N2X2 analogs.

X₁ = O or S

X₄ = O or S

A variant of the dichloroethane approach, shown above, can also lead to successful syntheses of this class of compounds. Reaction of an aminoalcohol or aminothiol derivative readily available from a natural or unnatural amino acid with an O-protected 1-chloro, 2-hydroxy ethane derivative followed by deprotection and substitution with chloride gives the mono-chloro compound shown which can be further reacted with an appropriate aminoalcohol or aminothiol derivative readily available from a natural or unnatural amino acid to give the un-symmetrical product shown. Deprotection followed by cyclization with a dichloroethane derivative would give a mixture of the the two position isomers shown.

2N2X series 2: when X₁ and X₃ are N and X₂ and X₄ are O or S then: R₃ and R₅ do not exist

R₂ and R₄ are independently chosen from H, CH₃, C2-C10 straight chain or branched alkyl, C3-C10, cycloalkyl, C1-C6 alkyl C3-C10 cycloalkyl, aryl, mono, di, tri, tetra and penta substituted aryl, heteroaryl, fused aryl, C1-C6 alkyl aryl, C1-C6 alkyl mono, di, tri, tetra and penta substituted aryl, C1-C5 alkyl heteroaryl, C1-C6 alkyl fused aryl, CH₂COOH, CH₂SO₃H, CH₂2PO(OH)₂,

CH₂P(CH₃O(OH); nl, n2, n3, and n4 are independently chosen to be 2 or 3, and each repeat of any of nl, n2, n3 and n4 may be the same as or different than any other repeat; and

R₇, R₈, R₉, R_1O, Rn, Ri₂, R₁₃ and Ru are independently chosen from H,

CH₃, C2-C10 straight chain or branched alkyl, C3-C10 cycloalkyl, C1-C6 alkyl

In addition, one or both of R₂, or R₄ may be functionalized for attachment, for example, to peptides, proteins, polyethylene glycols and other such chemical entities in order to modify the overall pharmacokinetics, deliverability and/or half lives of the constructs. Examples of such functionalization include but are not limited to Cl-ClO alkyl-CO-peptide, Cl-ClO alkyl-CO-protein, Cl-ClO alkyl-CO-PEG, Cl-ClO alkyl-NH-peptide, Cl-ClO alkyl-NH-protein, Cl-ClO alkyl-NH-CO-PEG, Cl-ClO alkyl-S-peptide, Cl-ClO alkyl-S-protein.

Furthermore one or several of R₇, R₈, R₉, Ri₀, Rn, Ri₂, Ri₃ or R₁₄ may be functionalized for attachment, for example, to peptides, proteins, polyethylene glycols and other such chemical entities in order to modify the overall pharmacokinetics, deliverability and/or half lives of the constructs. Examples of such functionalization include but are not limited to Cl-ClO alkyl-CO-peptide, Cl- ClO alkyl-CO-protein, Cl-ClO alkyl-CO-PEG, Cl-ClO alkyl-NH-peptide, Cl- ClO alkyl-NH-protein, Cl-ClO alkyl-NH-CO-PEG, Cl-ClO alkyl-S-peptide, and Cl-ClO alkyl-S-protein.

Synthesis of examples of the macrocyclic 2N2X series 2 of Formula I:

Triethylenetetramine itself has been synthesized by reaction of 2 equivalents of ethylene diamine with 1,2-dichloro ethane to give triethylenetetramine directly (1). Possible side products from this synthesis include the 12N4 macrocycle shown below, which could also be synthesized directly from Triethylenetetramine by reaction with a further equivalent of 1,2-dichloro ethane under appropriately dilute concentrations to provide the 12N4 macrocycle shown. Modification of this procedure by using starting materials with appropriate R groups would lead to symmetrically substituted 12N4 macrocycle examples as shown below:

X₂ = o or S

X4 = o or S

The judicious use of protecting group chemistry such as the widely used BOC (t-butyloxycarbonyl) group and an appropriate O or S protecting group allows the chemistry to be directed specifically towards the substitution pattern shown. Other approaches such as via the chemistry of ethyleneimine (2) may also lead to a subset of the di-aza 2X series. A variant of this approach using substituted dichloroethane derivatives could be used to access more complex substitution patterns. This would lead to mixtures of position isomers, which can be separated by HPLC. P

2 position isomers

4 position isomers

1N3X series: when X₁ is N and X₂, X₃ and X₄ are O or S then:

R₃, R₄ and R₅ do not exist;

R₂ is independently chosen from H₅ CH₃, C2-C10 straight chain or branched alkyl, C3-C10 cycloalkyl, C1-C6 alkyl C3-C10 cycloalkyl, aryl, mono, di, tri, tetra and penta substituted aryl, heteroaryl, fused aryl, C1-C6 alkyl aryl, C1-C6 alkyl mono, di, tri, tetra and penta substituted aryl, C1-C5 alkyl heteroaryl, C1-C6 alkyl fused aryl, CH₂COOH, CH₂SO₃H, CH₂PO(OH)₂, CH₂P(CH₃)O(OH); nl, n2, n3, and n4 are independently chosen to be 2 or 3, and each repeat of any of nl, n2, n3 and n4 may be the same as or different than any other repeat; and

R₇, R₈, R₉, R₁₀, R₁₁, R₁₂, Ri₃ and Ri₄ are independently chosen from H,

CH₃, C2-C10 straight chain or branched alkyl, C3-C10 cycloalkyl, C1-C6 alkyl

C3-C10 cycloalkyl, aryl, mono, di, tri, tetra and penta substituted aryl, heteroaryl, fused aryl, C1-C6 alkyl aryl, C1-C6 alkyl mono, di, tri, tetra and penta substituted aryl, C1-C5 alkyl heteroaryl, C1-C6 alkyl fused aryl.

In addition, R₂ may be functionalized for attachment, for example, to peptides, proteins, polyethylene glycols and other such chemical entities in order to modify the overall pharmaco-kinetics, deliverability and/or half lives of the constructs. Examples of such functionalization include but are not limited to Cl- ClO alkyl-CO-peptide, Cl-ClO alkyl-CO-protein, Cl-ClO alkyl-CO-PEG, Cl- ClO alkyl-NH-peptide, Cl-ClO alkyl-NH-protein, Cl-ClO alkyl-NH-CO-PEG, Cl-ClO alkyl-S-peptide, and Cl-ClO alkyl-S-protein.

Furthermore one or several Of R₇, Rg, R₉, Rio, Rn, Ri₂, Ri₃ or R₁₄ may be functionalized for attachment, for example, to peptides, proteins, polyethylene glycols and other such chemical entities in order to modify the overall pharmaco- kinetics, deliverability and/or half lives of the constructs. Examples of such functionalization include but are not limited to Cl-ClO alkyl-CO-peptide, Cl- ClO alkyl-CO-protein, Cl-ClO alkyl-CO-PEG, Cl-ClO alkyl-NH-peptide, Cl- ClO alkyl-NH-protein, Cl-ClO alkyl-NH-CO-PEG, Cl-ClO alkyl-S-peptide, and Cl-ClO alkyl-S-protein.

Synthesis of examples of the macrocyclic 1N3X series of Formula I:

(1). Possible side products from this synthesis include the 12N4 macrocycle shown below, which could also be synthesized directly from Triethylenetetramine by reaction with a further equivalent of 1,2-dichloro ethane under appropriately dilute concentrations to provide the 12N4 macrocycle shown. Modification of this procedure by using starting materials with appropriate R groups would lead to substituted 12NX3 macrocycle examples as shown below:

2 equivs

X2. ^X3. X4 = O or S

The judicious use of protecting group chemistry such as the widely used

BOC (t-butyloxycarbonyl) group and an appropriate O or S protecting group allows the chemistry to be directed specifically towards the substitution pattern shown. Other approaches such as via the chemistry of ethyleneimine (2) may also lead to a subset of the mono-aza 3X series. A variant of this approach using substituted dichloroethane derivatives could be used to access more complex substitution patterns. This would lead to mixtures of position isomers, which can be separated by HPLC.

2 position isomers

4 position isomers

For the tri-heteroatom acyclic examples of Formula II:

Xi, X₂, and X₃ are independently chosen from the atoms N, S or O such that: 3N series: when Xi, X₂, and X₃ are N then:

R₁, R₂, R₃, R₅, and R₆ are independently chosen from H, CH₃, C2-C10 straight chain or branched alkyl, C3-C10 cycloalkyl, C1-C6 alkyl C3-C10 cycloalkyl, aryl, mono, di, tri, tetra and penta substituted aryl, heteroaryl, fused aryl, C1-C6 alkyl aryl, C1-C6 alkyl mono, di, tri, tetra and penta substituted aryl, C1-C5 alkyl heteroaryl, C1-C6 alkyl fused aryl, CH₂COOH, CH₂SO₃H, CH₂PO(OH)₂, CH₂P(CH₃)O(OH); nl and n2 are independently chosen to be 2 or 3, and each repeat of any of nl and n2 may be the same as or different than any other repeat; and

R₇, R₈, R₉, and Ri₀ are independently chosen from H, CH₃, C2-C10 straight chain or branched alkyl, C3-C10 cycloalkyl, C1-C6 alkyl C3-C10 cycloalkyl, aryl, mono, di, tri, tetra and penta substituted aryl, heteroaryl, fused aryl, C1-C6 alkyl aryl, C1-C6 alkyl mono, di, tri, tetra and penta substituted aryl, C 1 -C5 alkyl heteroaryl, C 1 -C6 alkyl fused aryl.

. In addition, one or several of Ri, R₂, R₃, R₅ or R₆ may be functionalized for attachment, for example, to peptides, proteins, polyethylene glycols and other such chemical entities in order to modify the overall pharmaco-kinetics, deliverability and/or half lives of the constructs. Examples of such functionalization include but are not limited to Cl-ClO alkyl-CO-peptide, Cl- ClO alkyl-CO-protein, Cl-ClO alkyl-CO-PEG, Cl-ClO alkyl-NH-peptide, Cl- ClO alkyl-NH-protein, Cl-ClO alkyl-NH-CO-PEG, Cl-ClO alkyl- S-peptide, Cl- ClO alkyl-S-protein.

Furthermore one or several of R₇, Rg, R₉, or R^ may be functionalized for attachment, for example, to peptides, proteins, polyethylene glycols and other such chemical entities in order to modify the overall pharmaco-kinetics, deliverability and/or half lives of the constructs. Examples of such functionalization include but are not limited to Cl-ClO alkyl-CO-peptide, Cl-

ClO alkyl-CO-protein, Cl-ClO alkyl-CO-PEG, Cl-ClO alkyl-NH-peptide, Cl- ClO alkyl-NH-protein, Cl-ClO alkyl-NH-CO-PEG, Cl-ClO alkyl-S-peptide, and

Cl-ClO alkyl-S-protein. Synthesis of the open chain 3N series of Formula II:

As mentioned above Triethylenetetramine itself has been synthesized by reaction of 2 equivalents of ethylene diamine with 1,2-dichloro ethane to give Triethylenetetramine directly (1). A variant of this procedure by using starting materials with appropriate R groups and l-amino,2-chloro ethane would lead to some open chain 3N examples as shown below:

Trientine

The judicious use of protecting group chemistry such as the widely used

BOC (t-butyloxycarbonyl) group allows the chemistry to be directed specifically towards the substitution pattern shown. Other approaches such as via the chemistry of ethyleneimine (2) may also lead to a subset of the tri-aza series. In order to obtain the un-symmetrically substituted derivatives a variant of some chemistry described by Meares et al (2) could be used. Standard peptide synthesis using the Rink resin along with FMOC protected natural and un-natural amino acids which can be conveniently cleaved at the penultimate step of the synthesis generates a di-peptide C-terminal amide. This can be reduced using Diborane in THF to give the open chain tri-aza compounds as shown below:

^BHsinTHF *-

The reverse Rink approach may also be useful where peptide coupling is slowed for a particular substitution pattern as shown below. Again the incorporation of Ri , R₂, R₅ and R₆ can be accomplished with this chemistry by standard procedures:

_FMOCr^{H RθSin} — ^Resin

2NX series 1: when X₁ and X₃ are N and X₂ is S or O then: R₃ does not exist Ri, R₂, R₅, and R₆ are independently chosen from H, CH₃, C2-C10 straight chain or branched alkyl, C3-C10 cycloalkyl, C1-C6 alkyl C3-C10 cycloalkyl, aryl, mono, di, tri, tetra and penta substituted aryl, heteroaryl, fused aryl, C1-C6 alkyl aryl, C1-C6 alkyl mono, di, tri, tetra and penta substituted aryl, C1-C5 alkyl heteroaryl, C1-C6 alkyl fused aryl, CH₂COOH, CH₂SO₃H, CH₂PO(OH)₂, CH₂P(CH₃)O(OH); nl and n2 are independently chosen to be 2 or 3, and each repeat of any of nl and n2 may be the same as or different than any other repeat; and

R₇, R₈, R₉, and R₁₀ are independently chosen from H, CH₃, C2-C10 straight chain or branched alkyl, C3-C10 cycloalkyl, C1-C6 alkyl C3-C10 cycloalkyl, aryl, mono, di, tri, tetra and penta substituted aryl, heteroaryl, fused aryl, C1-C6 alkyl aryl, C1-C6 alkyl mono, di, tri, tetra and penta substituted aryl,

C1-C5 alkyl heteroaryl, C1-C6 alkyl fused aryl In addition, one or several of R₁, R₂, R₅ or R₆ may be functionalized for attachment, for example, to peptides, proteins, polyethylene glycols and other such chemical entities in order to modify the overall pharmaco-kinetics, deliverability and/or half lives of the constructs. Examples of such functionalization include but are not limited to Cl-ClO alkyl-CO-peptide, Cl- ClO alkyl-CO-protein, Cl-ClO alkyl-CO-PEG, Cl-ClO alkyl-NH-peptide, Cl- ClO alkyl-NH-protein, Cl-ClO alkyl-NH-CO-PEG, Cl-ClO alkyl-S-peptide, and Cl-ClO alkyl-S-protein.

Furthermore one or several of R₇, R₈, R₉, or R₁₀ may be functionalized for attachment, for example, to peptides, proteins, polyethylene glycols and other such chemical entities in order to modify the overall pharmaco-kinetics, deliverability and/or half lives of the constructs. Examples of such functionalization include but are not limited to Cl-ClO alkyl-CO-peptide, Cl- ClO alkyl-CO-protein, Cl-ClO alkyl-CO-PEG, Cl-ClO alkyl-NH-peptide, Cl- ClO alkyl-NH-protein, Cl-ClO alkyl-NH-CO-PEG, Cl-ClO alkyl-S-peptide, and Cl-ClO alkyl-S-protein.

Synthesis of the open chain 2NX series 1 of Formula II:

The synthesis of the 2NX series 1 compounds can be readily achieved as shown above. The judicious use of protecting group chemistry such as the widely used BOC (t-butyloxycarbonyl) group allows the chemistry to be directed specifically towards the substitution pattern shown above. Other approaches such as via the chemistry of ethyleneimine (2) may also lead to a subset of the tri-aza X series. 2NX series 2 when Xi and X₂ are N and X₃ is O or S then: R.₅ does not exist;

Ri, R₂, R₃ and R₆ are independently chosen from H, CH₃, C2-C10 straight chain or branched alkyl, C3-C10 cycloalkyl, C1-C6 alkyl C3-

ClO cycloalkyl, aryl, mono, di, tri, tetra and penta substituted aryl, heteroaryl, fused aryl, C1-C6 alkyl aryl, C1-C6 alkyl mono, di, tri, tetra and penta substituted aryl, C1-C5 alkyl heteroaryl, C1-C6 alkyl fused aryl, CH₂COOH, CH₂SO₃H, CH₂PO(OH)₂, CH₂P(CH₃)O(OH); nl and n2 are independently chosen to be 2 or 3, and each repeat of any of nl and n2 may be the same as or different than any other repeat; and

R₇, R₈, R₉, and Ri₀ are independently chosen from H, CH₃, C2-C10 straight chain or branched alkyl, C3-C10 cycloalkyl, C1-C6 alkyl C3- ClO cycloalkyl, aryl, mono, di, tri, tetra and penta substituted aryl, heteroaryl, fused aryl, C1-C6 alkyl aryl, C1-C6 alkyl mono, di, tri, tetra and penta substituted aryl, C1-C5 alkyl heteroaryl, C1-C6 alkyl fused aryl.

In addition, one or several of Ri, R₂, R₅, or R₆ may be functionalized for attachment, for example, to peptides, proteins, polyethylene glycols and other such chemical entities in order to modify the overall pharmaco-kinetics, deliverability and/or half lives of the constructs. Examples of such functionalization include but are not limited to Cl-ClO alkyl-CO-peptide, Cl-

Cl-ClO alkyl-S-protein.

Synthesis of the open chain 2NX series 2 of Formula II:

^Resin →

S

For the cases where X₃ = O or S a similar approach using standard peptide synthesis according to the Rink approach as shown above can be used. Coupling of a suitably protected alpha thiolo or hydroxy carboxylic acid with a Rink resin amino acid derivative followed by cleavage gives the desired linear di-amide, which can be reduced with Diborane in THF to give the open chain 2NX compounds. The incorporation of R₁, R₂, R₅ and R₆ can be accomplished with this chemistry by standard procedures.

The reverse Rink version is also feasible and again the incorporation of R₁, R₂, R₅ and R₆ can be accomplished with this chemistry by standard procedures.

Resin

S

Tri-heteroatom cyclic series of Formula II:

R₁ and R₆ form a bridging group (CR_πR₁₂)n3; and Xi, X₂, and X₃ are independently chosen from the atoms N, S or O such that: 3N series: when Xi, X₂ and X₃ are N then:

R₂, R₃, and R₅ are independently chosen from H, CH₃, C2-C10 straight chain or branched alkyl, C3-C10 cycloalkyl, C1-C6 alkyl C3-C10 cycloalkyl, aryl, mono, di, tri, tetra and penta substituted aryl, heteroaryl, fused aryl, C1-C6 alkyl aryl, C1-C6 alkyl mono, di, tri, tetra and penta substituted aryl, C1-C5 alkyl heteroaryl, C1-C6 alkyl fused aryl, CH₂COOH, CH₂SO₃H, CH₂PO(OH)₂, CH₂P(CH₃)O(OH); nl, n2, and n3 are independently chosen to be 2 or 3, and each repeat of any of nl, n2 and n3 may be the same as or different than any other repeat; and

R₇, R₈, R₉, Rio, Rn, and R₁₂ are independently chosen from H, CH₃,

C2-C10 straight chain or branched alkyl, C3-C10 cycloalkyl, C1-C6 alkyl C3-

In addition, one or several of R₂, R₃, or R₅ may be functionalized for attachment, for example, to peptides, proteins, polyethylene glycols and other such chemical entities in order to modify the overall pharmaco-kinetics, deliverability and/or half lives of the constructs. Examples of such functionalization include but are not limited to Cl-ClO alkyl-CO-peptide, Cl- ClO alkyl-CO-protein, Cl-ClO alkyl-CO-PEG, Cl-ClO alkyl-NH-peptide, Cl- ClO alkyl-NH-protein, Cl-ClO alkyl-NH-CO-PEG, Cl-ClO alkyl- S-peptide, and Cl-ClO alkyl-S-protein.

Furthermore one or several of R₇, R₈, R₉, Rio, Rn, or Ri₂ may be functionalized for attachment, for example, to peptides, proteins, polyethylene glycols and other such chemical entities in order to modify the overall pharmaco- kinetics, deliverability and/or half lives of the constructs. Examples of such functionalization include but are not limited to Cl-ClO alkyl-CO-peptide, Cl- ClO alkyl-CO-protein, Cl-ClO alkyl-CO-PEG, Cl-ClO alkyl-NH-peptide, Cl- ClO alkyl-NH-protein, Cl-ClO alkyl-NH-CO-PEG, Cl-ClO alkyl-S-peptide, Cl- ClO alkyl-S-protein.

Synthesis of examples of the macrocyclic 3N series of Formula II: As mentioned above Triethylenetetramine itself has been synthesized by reaction of 2 equivalents of ethylene diamine with 1,2-dichloro ethane to give Triethylenetetramine directly (1). A variant of this procedure by using starting materials with appropriate R groups and l-amino,2-chloro ethane would lead to open chain 3N examples which could then be cyclized by reaction with an appropriate 1,2 dichloroethane derivative as shown below:

_H2N^NH_{2 + ci} CI ^ H₂N^ N^^NH2

H Trientine

The judicious use of protecting group chemistry such as the widely used

BOC (t-butyloxycarbonyl) group allows the chemistry to be directed specifically towards the substitution pattern shown. Other approaches such as via the chemistry of ethyleneimine (2) may also lead to a subset of the macrocyclic tri- aza series. In order to obtain the un-symmetrically substituted derivatives a variant of some chemistry described by Meares et al (3) could be used. Standard peptide synthesis using the Merrifield approach/SASRIN resin along with FMOC protected natural and un-natural amino acids which can be conveniently cleaved at the penultimate step of the synthesis generates a tri-peptide attached to resin via its C-terminus. This can be cyclized during concomitant cleavage from the resin followed by reduction using Diborane in THF to give the cyclic tri-aza compounds as shown below:

^BH3inτHF _>

The incorporation of R₁, R₂, and R₅ can be accomplished with this chemistry by standard procedures. The reverse Rink approach may also be useful where peptide coupling is slowed for a particular substitution pattern as shown below. Again the incorporation of R₁, R₂, R₅ and R₆ can be accomplished with this chemistry by standard procedures:

2NX series: when X₁ and X₂ are N and X₃ is S or O then: R₅ does not exist;

R₂ and R₃ are independently chosen from H, CH₃, C2-C10 straight chain or branched alkyl, C3-C10 cycloalkyl, C1-C6 alkyl C3-C10 cycloalkyl, aryl, mono, di, tri, tetra and penta substituted aryl, heteroaryl, fused aryl, C1-C6 alkyl aryl, C1-C6 alkyl mono, di, tri, tetra and penta substituted aryl, C1-C5 alkyl heteroaryl, C1-C6 alkyl fused aryl, CH₂COOH, CH₂SO₃H, CH₂PO(OH) ₂, CH₂P(CH₃)O(OH); nl, n2, and n3 are independently chosen to be 2 or 3, and each repeat of any of nl, n2 and n3 may be the same as or different than any other repeat; and R₇, R₈, R₉, R_1O, Rn, and R₁₂ are independently chosen from H, CH₃,

C2-C10 straight chain or branched alkyl, C3-C10 cycloalkyl, C1-C6 alkyl C3- ClO cycloalkyl, aryl, mono, di, tri, tetra and penta substituted aryl, heteroaryl, fused aryl, C1-C6 alkyl aryl, C1-C6 alkyl mono, di, tri, tetra and penta substituted aryl, C1-C5 alkyl heteroaryl, C1-C6 alkyl fused aryl. In addition, one or both of R₂ or R₃ may be functionalized for attachment, for example, to peptides, proteins, polyethylene glycols and other such chemical entities in order to modify the overall pharmacokinetics, deliverability and/or half lives of the constructs. Examples of such functionalization include but are not limited to Cl-ClO alkyl-CO-peptide, Cl-ClO alkyl-CO-ρrotein, Cl-ClO alkyl-CO-PEG, Cl-ClO alkyl-NH-peptide, Cl-ClO alkyl-NH-protein, Cl-ClO alkyl-NH-CO-PEG, Cl-ClO alkyl-S-peptide, and C 1 -C 10 alkyl-S-protein.

Furthermore one or several of R₇, R₈, R₉, Rio, Rn, or R_]2 may be functionalized for attachment, for example, to peptides, proteins, polyethylene glycols and other such chemical entities in order to modify the overall pharmacokinetics, deliverability and/or half lives of the constructs. Examples of such functionalization include but are not limited to Cl-ClO alkyl-CO-peptide, Cl- ClO alkyl-CO-protein, Cl-ClO alkyl-CO-PEG, Cl-ClO alkyl-NH-peptide, Cl- ClO alkyl-NH-protein, Cl-ClO alkyl-NH-CO-PEG, Cl-ClO alkyl-S-peptide, and Cl-ClO alkyl-S-protein.

Synthesis of examples of the macrocyclic 2NX series of Formula II:

As mentioned above Triethylenetetramine itself has been synthesized by reaction of 2 equivalents of ethylene diamine with 1,2-dichloro ethane to give Triethylenetetramine directly (1). A variant of this procedure by using starting materials with appropriate R groups and l-amino,2-chloro ethane would lead to open chain 2NX examples which could then be cyclized by reaction with an appropriate 1,2 dichloroethane derivative as shown below:

Trientine

X₃ = S or O

The judicious use of protecting group chemistry such as the widely used BOC (t-butyloxycarbonyl) group allows the chemistry to be directed specifically towards the substitution pattern shown. Other approaches such as via the chemistry of ethyleneimine (2) may also lead to a subset of the macrocyclic di- aza X series. In order to obtain the un-symmetrically substituted derivatives a variant of some chemistry described by Meares et al (3) could be used. Standard peptide synthesis using the Merrifield approach/SASRIN resin along with FMOC protected natural and un-natural amino acids which can be conveniently cleaved at the penultimate step of the synthesis generates a tri-peptide attached to resin via its C-terminus. This can be cyclized during concomitant cleavage from the resin followed by reduction using Diborane in THF to give the cyclic tri-aza compounds as shown below:

The incorporation OfR₁, and R₂ can be accomplished with this chemistry by standard procedures.

The reverse Rink approach may also be useful where peptide coupling is slowed for a particular substitution pattern as shown below. Again the incorporation Of R₁, and R₂ can be accomplished with this chemistry by standard procedures:

BH₃ Jn THF Tosylation *^•

S

1N2X series: when X₁ is N and X₂ and X₃ are O or S then:

R₃ and R₅ do not exist;

R₂ is independently chosen from H, CH₃, C2-C10 straight chain or branched alkyl, C3-C10 cycloalkyl, C1-C6 alkyl C3-C10 cycloalkyl, aryl, mono, di, tri, tetra and penta substituted aryl, heteroaryl, fused aryl, C1-C6 alkyl aryl, C1-C6 alkyl mono, di, tri, tetra and penta substituted aryl, C1-C5 alkyl heteroaryl, C1-C6 alkyl fused aryl, CH₂COOH, CH₂SO₃H, CH₂PO(OH)₂, CH₂P(CH₃)O(OH); nl, n2, and n3 are independently chosen to be 2 or 3, and each repeat of any of nl, n2 and n3 may be the same as or different than any other repeat;

R₇, Rg, R₉, Rio, Rn, and Ri₂ are independently chosen from H, CH₃,

C2-C10 straight chain or branched alkyl, C3-C10 cycloalkyl, C1-C6 alkyl C3-

Furthermore one or several of R₇, R₈, R₉, Rio, Rn, or R_n may be functionalized for attachment, for example, to peptides, proteins, polyethylene glycols and other such chemical entities in order to modify the overall pharmacokinetics, deliverability and/or half lives of the constructs. Examples of such functionalization include but are not limited to Cl-ClO alkyl-CO-peptide, Cl- ClO alkyl-CO-protein, Cl-ClO alkyl-CO-PEG, Cl-ClO alkyl-NH-peptide,' Cl- ClO alkyl-NH-protein, Cl-ClO alkyl-NH-CO-PEG, Cl-ClO alkyl-S-peptide, and Cl-ClO alkyl-S-protein. Synthesis of examples of the macrocyclic 1N2X series of Formula II:

As mentioned above Triethylenetetramine itself has been synthesized by reaction of 2 equivalents of ethylene diamine with 1,2-dichloro ethane to give Triethylenetetramine directly (1). A variant of this procedure by using starting materials with appropriate R groups and l-amino,2-chloro ethane would lead to open chain 1N2X examples which could then be cyclized by reaction with an appropriate 1,2 dichloroethane derivative as shown below:

Trientine

R₉ R₇ R9 Rδ

.NH₂

Cl' ^~~*~ PrOtX₃" A . X₂^

R-io Rf ! R-io R7

^X2, X3 = S or O

The judicious use of protecting group chemistry such as the widely used BOC (t-butyloxycarbonyl) group allows the chemistry to be directed specifically towards the substitution pattern shown. Other approaches such as via the chemistry of ethyleneimine (2) may also lead to a subset of the macrocyclic aza di-X series. In order to obtain the un- symmetrically substituted derivatives a variant of some chemistry above could be used: ^! H₂

X₂, X₃ = S or O

The incorporation of R₁ and R₂ can by accomplished with this chemistry by standard procedures.

Copper antagonists and pharmaceutically acceptable salts for use according to the present invention may also be synthesized using methods described in U.S. Patent Application No. 11/184,761, filed 07/19/2005, the contents of which are hereby incorporated by reference in its entirety. Medical devices of the invention may include any device used or implanted in or on a mammal, including a human or an animal host. Examples include stents, balloons, prosthetic heart valves, annuloplasty rings, ventricular assist devices, including left ventricular assist devices, right ventricular assist devices, and biventricular assist devices, grafts, shunts, sewing rings (including those having silicone or polyurethane inserts), polyester fabric encasements, medical leads, orthopedic plates, bone pins, bone substitutes, anchors, joints, screws, ophthalmic implants (including, for example, orbital implants, lens implants, corneal implants (including intrasomal corneal ring segments (INTACS)), and microchips), catheters, cannulae, pulse generators, cardiac defibrillators, arteriovenous shunts, pacemakers, sutures, suture anchors, staples, anastomosis devices, vertebral disks, hemostatic barriers, clamps, clips, vascular implants, tissue adhesives and sealants, tissue scaffolds, various types of dressings, intraluminal devices, and vascular supports.

Any one or more of the foregoing medical devices can include an overlayer of any type, including, for example, a fabric such as a sheath, an encasement, a layer, or a coating, such that the fabric overlayer is in contact with body tissue or fluids such as blood. Alternatively, instead of a fabric overlayer, the medical device may include any other type of layer such as, for example, a mesh, coil, wire, inflatable balloon, bead, sheet, or any other structure which is capable of being used or implanted at a target location, including, for example, intravascular target locations, intraluminal target locations, intra-orbital target locations, target locations within solid tissue, eyes, etc.

Exemplary medical devices that are intended as tissue implants include, for example, brachytherapy sources, embolization materials, tumor-bed implants, intra-joint implants, materials to minimize adhesions, etc. Examples of stents include intravascular and intraductal stents, and see also, for example, Pepine et al., "Coronary Artery Stents, JACC Vol. 28, No. 3, September 1996:782-94; D. Stoeckel, "A survey of stent designs," Min Invas Ther & Allied Technol 2002: 11(4) 137-147. Stents may also include, for example, balloon-expandable stents and self-expanding stents. Balloon-expandable stents include those of the sort available from a number of commercial suppliers, including Cordis Johnson & Johnson Interventional Systems, Medi-Tech, Cook, ACS, and Metronic.

Self-expanding stents are typically composed, for example, from a shape memory alloy and are available from suppliers, such as Instent. In the case of stents, a balloon-expandable stent is typically composed of a stainless steel framework or, in the case of self-expanding stents, from nickel/titanium alloy. While typically stents are made of a metallic material for strength purposes, polymeric or plastic materials may also be utilized in the ^stent construction. While it is preferred that the stent be coated in an expanded position, coating the stent in an unexpanded position is also contemplated. Exemplary coated balloons include, for example, coated balloons and coated balloon catheters, including inflatable and self inflatable coated balloons and balloon catheters. The inflatable coated balloon may be a non-dispensable balloon, for example, typically composed of polyethyleneterephthalate, or it may be an elastic balloon, for example, typically being composed of latex or silicone rubber.

Delivery of copper antagonists utilizing a stent can be carried out in a number of ways, including, for example, from the struts of a stent, a stent graft, the catheter used to deliver the stent, the stent cover or sheath. Various methods of applying a therapeutic agent to a stent and administering the therapeutic agent via the stent are disclosed in U.S. Patent Nos. 6,702,850; 6,585,764; 6,358,556; 6,344,028; 6,251,136; 5,697,967; 5,599,352; 5,591,227; 5,464,650; 5,304,121; 5,163,952; 5,092,877; 4,994,071; and 4,916,193, the disclosures of which are incorporated in their entirety herein by reference. Other devices may be coated or impregnated using these or other techniques in the art, including those now known or later developed.

There are various techniques known in the art for the localized delivery of therapeutic agents, for example, but not limited to, by means of a small catheter which extends from the exterior of the subject to the internal tissue site, with a mechanical delivery system being provided to administer a therapeutic agent in a continuous or periodic controlled dosage. This method can be utilized for subjects in need of therapeutic treatment for a substantial period of time.

Any catheter, however, is within the scope of the invention, including those for the delivery of therapeutic agents or otherwise. These include, for example, urological catheters (including intermittent catheters, external catheters, and

Foley catheters), pancreatic catheters, hepatic catheters, infusion catheters, cardiovascular catheters, renal catheters, hemodynamic monitoring catheters, neurological catheters, and so on. Cannulae, as noted above, are also included.

The method of applying one or more copper antagonists to an internal tissue site of a subject comprises, for example, advancing an elongate member, such as a catheter (which may itself be coated or impregnated with one or more copper antagonists), internally into the subject to cause a portion of the elongated member to occupy the internal tissue site. A portion of the elongated member comprises a lateral wall section which carries the copper antagonist in a manner permitting release thereof from the lateral wall section at the internal tissue site once the site has been reached. This can be determined by the usual methods, such as for example, fibre optic television, x-ray, etc.

The release of the copper antagonist at the internal tissue site can occur for example by the use of a catheter balloon, which when inflated causes the copper antagonist to be pressed into and/or onto the tissue at the internal tissue site. This allows at least some of the copper antagonist to be retained at the tissue site once the catheter balloon is deflated.

The copper antagonist can also be mixed with a controlled release carrier and administered in the manner discussed above, for example. Such controlled release carriers can be biodegradable over a relatively long period of time, for example, over a period of days, weeks or even months, so that as the controlled release carriers are brought into contact with the tissues and/or fluids the controlled release carriers degrade over time to allow for a relatively slow, controlled diffusion of the copper antagonist to the tissue and/or fluids. The carrier over time will be removed by natural bodily processes. The catheter or balloon catheter or other device used to apply a stent to the coronary artery or elsewhere may also be provided with a coating of heparin or other anti-thrombogenic agent, for example, in conjunction with one or more copper antagonists that may be combined with a carrier or a controlled release carrier for the copper antagonist. Thus, simultaneously with the application of the copper antagonist, the heparin or other anti-thrombogenic agent, for example, is applied to the internal tissue site for the long term suppression of thrombogenic activity in the vicinity of the stent in addition to therapeutic effects from the copper antagonist. ^!

In addition, while the catheter or other elongate member, for example, is being used to position the stent and is being advanced to the internal tissue site, the portion of the catheter carrying the copper antagonist may be enclosed in a protective sheath. The sheath is used to prevent removal of substantial amounts of the copper antagonist from the catheter before reaching the desired internal site. When the site is reached, the protective sheath may be withdrawn to expose the catheter portion carrying the copper antagonist. The copper antagonist can then be applied to the internal site, for example, by expansion of a catheter balloon upon which the copper antagonist resides, or by other processes such as spontaneous dispersion off the catheter into the tissues. If desired, the protective sheath may be a conventional introducer catheter, or it may be a split introducer sheath to facilitate removal of the sheath from the catheter after its withdrawal, for example.

One example of a method of coating a medical device having more than one surface or requiring only a portion of a surface of the medical device to be treated is to treat the device with gas plasma that may, for example, be composed of a molecular species containing the copper antagonist. In the case of stents, it is particularly desirable to treat the entire surface. In the case of balloons mounted on catheters, it is desirable to coat at least the outer cylindrical surface of the balloon that will be in contact with a blood vessel or other tissue when the balloon is inflated.

Alternatively, the copper antagonist can be mixed with polymers (both degradable and non degradable), for example, to hold a copper chelator(s) to a stent or graft or other device, or the copper antagonist can be entrapped into the copper or plastic or other material of, for example, a stent or graft body.

Alternatively the copper antagonist can be covalently bound to, for example, a stent or other device via solution chemistry techniques or dry chemistry techniques (for example, vapour deposition methods such as rf-plasma polymerization) and combinations thereof.

Another method of coating a surface of a medical device with one or more copper antagonists comprises contacting the surface with a copper antagonist(s) so as to cause the surface to be coated with the particular copper antagonist. Coating of the artificial surface may be accomplished using the methods described in the Examples, or other standard methods well known to those of ordinary skill in the art.

For example, coating a surface with a copper antagonist can be achieved by bathing the artificial surface, either by itself or within a device, in a solution containing the copper antagonist. In addition, synthetic copper antagonists may be coated onto an artificial surface by a variety of chemical techniques which are well known in the art. Such techniques include attaching the copper antagonist, for example, a functionalized compound of Formula I or II herein, by means of a linking group, or to a nucleophilic center, copper, epoxide, lactone, an alpha- or beta-saturated carbon chain, alkyl halide, carbonyl group, or Schiff base, by way of a reactive group, for example, a free thiol.

For example, triethylenetetramine and other compounds, such as those described herein, for example, may be derivatized to include a thiol moiety to yield S-triethylenetetramine. S -triethylenetetramine may then be linked to a polymer containing at least one accessible sulphur atom such as, for example, a polypeptide comprising one or more accessible cysteine residues, the sulphur atom of which is able to form a disulfide bond with the sulphur of the S- triethylenetetramine, thereby covalently linking said S-triethylenetetramine with said polymer. A medical device may be coated using a variety of different techniques.

The coating may be applied as a mixture, solution or suspension of polymeric material, for example, and one or more finely divided copper antagonists dispersed in an organic vehicle or a solution or partial solution of such copper chelator(s) or binding compound(s) in a solvent or vehicle for the polymer and/or copper chelator(s) or binding compound(s). For the purposes of this patent, the term "finely divided" means any type or size of included material from dissolved molecules through suspensions, colloids and particulate mixtures, or that otherwise serve the intended purpose. One or more copper antagonists can be disbursed in the carrier material, which may be the polymer, a solvent, or both, for example. The coating may be applied as a single layer or as a plurality of layers, typically relatively thin layers, sequentially applied, for example, in relatively rapid sequence. In some applications, the coating may further be characterized as a composite initial tie coat, or undercoat, and a composite topcoat. The coating thickness ratio of the topcoat to the undercoat may vary with the desired effect and/or the elution system. Typically, the topcoat and undercoat are of different formulations, but need not be. Providing a copper chelator(s) and/or binding compound(s) as a plurality of layers on the medical device enables both an initial burst effect of drug elution and the drug release kinetic profile associated with long term therapeutic effect to be controlled.

Various combinations of coating materials, such as polymer coating materials, for example, can be coordinated with biologically or chemically active species of interest to produce desired effects when coated on stents or other medical devices to be implanted, or inserted, in accordance with the invention. Loadings of therapeutic materials may vary, as well as the types of therapeutic material. The mechanism of incorporation of the biologically or chemically active species into the surface coating, as well as the egress mechanism, depends both on the nature of the surface coating polymer and the therapeutic material to be incorporated. The mechanism of release also depends on the mode of incorporation. The therapeutic material may elute via interparticle paths or be administered via transport or diffusion through the encapsulating material itself. Suitable polymers for use in the coating include, for example, a polymer that is biocompatible and minimizes irritation to the vessel wall when a medical device is implanted. It is advantagous that such polymer exhibit high elasticity/ductility, resistance to erosion, elasticity, and controlled drug release. Such polymer may be either a biostable or a bioabsorbable polymer depending on the desired rate of release or the desired degree of polymer stability. Bioabsorbable polymers that may be used include poly(L-lactic acid), polycaprolactone, poly(lactide-co-glycolide), poly(ethylene-vinyl acetate), poly(hydroxybutyrate-co-valerate), '■ polydioxanone, polyorthoester, polyanhydride, poly(glycolic acid), poly(D,L-lactic acid), poly(glycolic acid-co- trimethylene carbonate), polyphosphoester, polyphosphoester urethane, poly(amino acids), cyanoacrylates, poly(trimethylene carbonate), poly(iminocarbonate), copoly(ether-esters) (e.g. PEO/PLA), polyalkylene oxalates, polyphosphazenes and biomolecules such as fibrin, fibrinogen, cellulose, starch, collagen and hyaluronic acid.

Also, biostable polymers with a relatively low chronic tissue response such as polyurethanes, silicones, and polyesters could be used and other polymers may also be used if they can be dissolved and cured or polymerized on the medical device such as polyolefins, polyisobutylene and ethylene-alphaolefin copolymers; acrylic polymers and copolymers, ethylene-co-vinylacetate, polybutylmethacrylate, vinyl halide polymers and copolymers, such as polyvinyl chloride; polyvinyl ethers, such as polyvinyl methyl ether; polyvinylidene halides, such as polyvinylidene fluoride and polyvinylidene chloride; polyacrylonitrile, polyvinyl ketones; polyvinyl aromatics, such as polystyrene, polyvinyl esters, such as polyvinyl acetate; copolymers of vinyl monomers with each other and olefins, such as ethylene-methyl methacrylate copolymers, acrylonitrile-styrene copolymers, ABS resins, and ethylene- vinyl acetate copolymers; polyamides, such as Nylon 66 and polycaprolactam; alkyd resins; polycarbonates; polyoxymethylenes; polyimides; polyethers; epoxy resins, polyurethanes; rayon; rayon-triacetate; cellulose, cellulose acetate, cellulose butyrate; cellulose acetate butyrate; cellophane; cellulose nitrate; cellulose propionate; cellulose ethers; and carboxymethyl cellulose.

Accordingly, the coating comprising a copper antagonist may be formed, at least in part, for example, from a biodegradable or bioresorbable polymer material. Polymer materials can include, for example, but are not limited to, nylon, polyethylene perthalate, polytetrafluoroethylene, etc. Other polymers may also include, for example, hexafluoropropylene (HFP), tetrafluoroethylene (TFE), vinylidenefluoride, 1-hydropentafluoropropylene, perfluoro(methyl vinyl ether), chlorotrifluoroethylene (CTFE), pentafluoropropene, trifluoroethylene, hexafluoroacetone and hexafluoroisobutylene.

The desired release rate profile can be tailored by, for example, varying the coating thickness, the radial distribution (layer to layer) of bioactive materials, the number of layers, the mixing method, and the amount of bioactive material(s), the combination of different materials, for example, matrix polymer materials, at different layers, and the crosslink density of a polymeric material. The crosslink density is related to the amount of crosslinking which takes place and also the relative tightness of the matrix created by the particular crosslinking agent used. Thus, the curing process of such a coating typically determines the amount of crosslinking and also the crosslink density of the polymer material. For bioactive materials released from a crosslinked matrix, a crosslink structure of greater density will increase release time and reduce burst effect. By applying at least one therapeutically active copper antagonist at the outer layer of the medical device a burst effect may be made to occur where a large amount of the copper antagonist is immediately or promptly released when it comes into contact with the tissue. Subsequently, longer term release of the copper antagonist will occur as it diffuses through the material, for example, a polymeric material. The elution kinetics of a copper antagonist can be modified to meet the needs of the particular medical device application. For example, medical devices can be coated using a combination of a copper antagonist with one, two or more other medicaments, where the release sequence can be rate controlled. For example, one or more copper antagonists may be combined in the undercoat layer, and anti-thrombotic drugs, for example, heparin, may be provided in the topcoat layer. In this manner, the anti-thrombotic drugs will elute first, followed by the copper antagonist(s). In the case where the medical device is an implanted stent, this combination of drugs may better enable safe encapsulation of the implanted stent. Any desired drug may be included in this manner. Alternately, the drag coating may include a base coat layer applied directly to the surfaces of the medical device, a second layer which includes a pharmacological agent, for example, a copper antagonist, and a third layer in the form of a continuous membrane encapsulating the entire device. The base coat serves as a primer by readily adhering to the surface of the medical device and then readily accepting and retaining the copper antagonist(s) applied thereto. The base coat may include materials such as vitronectin, fibronectin, gelatin, collagen, and/or other similar materials, for example, which are relatively inexpensive and dry to form a sticky coating. A copper antagonist may be supplied in the form of dry, micronized particles, for example, that readily adhere to the sticky base layer surface. It is preferred that the copper antagonist may have a particle size of about 0.005 to about 3.0 micro metres, or such size may be different, if desired. Other particle sizes are contemplated depending on the particular medical application and device to which the copper antagonist(s) is/are being applied. The outer membrane or layer may encapsulate the entire medical device to cover all of its surfaces, including any bare device structure, any exposed base coating or the layer of micronized copper antagonist(s) or other medicament particles.

The material selected to form the membrane is dependent on its membrane forming characteristics and its biocompatibility,^' as well as its permeability to a copper antagonist. The chemical composition of the membrane forming polymer, for example, and that of a copper antagonist, in combination with the thickness of the applied outer layer, will determine the diffusion rate of the copper antagonist.

The overall coating should be thin enough so that it will not significantly increase the profile of the medical device when inserted into a mammal. In the case of implantable stents, the coating is, for example, from between 0.005 microns to about 400 microns thick. However, other thicknesses may be utilized without departing from the spirit and scope of the present invention. The adhesion of the coating and the rate at which the drug is delivered can be controlled by the selection of an appropriate bioabsorbable or biostable material, such as a polymer, and by the ratio of the drug-to-polymer in the solution. In the case where multiple layers are utilized to coat the medical device, the release rate can be further controlled by varying the ratio of, for example, copper antagonist- to-polymer, in the multiple layers. For example, a higher copper antagonist-to- polymer ratio in the outer layers than in the inner layers would result in a higher early dose which would decrease over time. In an alternate form, the medical device can include reservoirs, or channels, which may be loaded with one or more copper antagonists. Such reservoirs can aid in decreasing the profile of the coated device, since the copper antagonist or a portion of the copper antagonist would be provided within the reservoir or channel. In such an embodiment, the copper antagonist is provided in the reservoirs, and a coating or membrane of biocompatible material is applied over the reservoir which controls the diffusion of the copper antagonist from the reservoirs to the tissue. Further layers of copper antagonist and/or materials, for example, polymeric materials, may be applied to the device in accordance with the teachings herein without departing from the spirit and scope of the present invention.

The amount of copper antagonist included in the layer(s), will vary depending on the dosage required for effective therapeutic treatment. A therapeutically effective amount of a copper chelator, for example, one or more triethylenetetramine active agents, including but not limited to triethylenetetramine, triethylenetetramine salts, triethylenetetramine analogs of Formulas I and II, and so on, may be determined from the doses of such compounds administered orally that typically will vary from about 100 mg to about 3600 mg per day, and are typically in the range of about 600 mg to about 1200 mg to about 2400 mg per day. Other therapeutically effective dose ranges include, for example, from about 20 mg to about 3.9 g, from about 30 mg to about 3.7 g, from about 40 mg to about 3.5 g, from about 50 mg to about 3 g, from about 60 mg to about 2.8 g, from about 70 mg to about 2.5 g, about 80 mg to about 2.3 g, about 100 mg to about 2 g, about 100 mg to about 1.5 g, about 200 mg to about 1400 mg, about 200 mg to about 1300 mg, about 200 mg to about 1200 mg, about 200 mg to about 1100 mg, about 200 mg to about 1000 mg, about 300 mg to about 900 mg, about 300 mg to about 800 mg, about 300 mg to about 700 mg or about 300 mg to about 600 mg per day.

The amount of copper antagonist in the coating or layer is adjusted so that the desired dose of copper antagonist is delivered at the desired delivery rate for the desired time of delivery. The time of delivery will depend on factors which include the time period for which the device is intended to be implanted in the subject.

In one embodiment the coating may contain copper antagonist in a weight percentage of from about 0.0001% to about 30%. As noted above, depending on dose, rate of delivery, period of delivery and other factors, other amounts are contemplated and may be used. Thus, according to one option, the coating may contain copper antagonist in a weight percentage range of about 0.001% to about 25%, alternatively in a range of about 0.01% to about 20%, about 0.1% to about 15%, about 0.5% to about 12%, about 1% to about 10%, about 2% to about 10%, about 5% to about 10%, about 0.01% to about 5%, about 0.1% to about 5% or about 0.5% to about 5%. The weight percentage for the copper antagonist will be adjusted as appropriate, in view of considerations which include, but are not limited to, the following: the dose of copper antagonist to be delivered locally, the rate of release of copper antagonist from the coating and the time period for delivery of copper antagonist.

Copper antagonists, including but not limited to triethylenetetramine active agents and compounds of Formula I and II, and the like, will also be effective at doses in the order of 1/10, 1/50, 1/100, 1/200, 1/300, 1/400, 1/500 and even 1/1000 of those described herein. For example, low dose copper antagonists may include compounds, including copper chelators, particularly Cu+2 chelators, including but not limited to triethylenetetramine active agents and compounds of Formula I and II, and the like, in an amount sufficient to provide, for example, dosages from about 0.001 mg/kg to about 5 mg/kg, about 0.01 mg/kg to about 4.5 mg/kg, about 0.02 mg/kg to about 4 mg/kg, about 0.02 mg/kg to about 3.5 mg/kg, about 0.02 mg/kg to about 3 mg/kg, about 0.05 mg/kg to about 2.5 mg/kg, about 0.05 mg/kg to about 2 mg/kg, about 0.05-0.1 mg/kg to about 5 mg/kg, about 0.05-0.1 mg/kg to about 4 mg/kg, about 0.05-0.1 mg/kg to about 3 mg/kg, about 0.05-0.1 mg/kg to about 2 mg/kg, about 0.05-0.1 mg/kg to about 1 mg/kg, and/or any other doses or dose ranges within the ranges set forth herein. Other therapeutically effective dosage ranges may be useful. One skilled in the art can customize the desired rate and/or dosage of copper antagonist delivery by evaluation and/or selection of an appropriate bioabsorbable or biostable polymer and by the ratio of copper antagonist-to-polymer in the coating, for example.

The particular surface or surfaces on which the copper antagonist is deposited determines where the copper antagonist will be delivered upon implantation. For example, in the case of a stent, copper antagonist deposited on the outer exterior surfaces of the stent will cause the copper antagonist to pass directly into the lumen wall, while deposition of the copper antagonist on the outer interior surfaces of the stent will cause the copper antagonist to be released directly into the blood stream. Alternately, coating only the upstream edge or only the downstream edge of the stent may be desirable to achieve a desired effect. By selectively coating the stent, or other medical device, surfaces with the copper antagonist or other medicaments, the distribution of the copper antagonist may be precisely controlled. It is also contemplated that artificial surfaces will vary depending on the nature of the surface, and such characteristics as contour, crystallinity, hydrophobicity, hydrophilicity, capacity for hydrogen bonding, and flexibility of the molecular backbone and polymers. Therefore, using routine methods, one of ordinary skill will be able to customize the coating technique by adjusting such parameters as the amount of copper antagonist, length of treatment, temperature, diluents, and storage conditions, in order to provide optimal coating of each particular type of surface.

After the device or artificial material has been coated or impregnated with one or more copper antagonists, it will be suitable for its intended use, for example, implantation as a heart valve, insertion as a catheter, or insertion as a stent, and so on. The coated device or artificial surface will be suitable for use in conjunction with an animal, generally mammals, including humans.

Another embodiment of a copper antagonist pertains to the derivatization of synthetically derived polymeric materials by attachment of a functionalized copper antagonist such as, for example, a functionalized compound of Formula I or II as described herein. In another embodiment the invention also relates to a method and product for administering one or more copper antagonists in combination with one or more therapeutic agents. Therapeutic agents may include, for example, anti- thrombogenic agents, anticoagulants, antiplatelet agents, thrombolytics, antiproliferatives, anti-inflammatory agents, statins, α-adrenergic receptor antagonists, β_rselective adrenergic antagonists, ACE inhibitors, calcium channel blockers, angiotensin II receptor antagonists, vasodilators, antiproliferative/antimitotic agents, immunosuppressive agent, agents that inhibit hyperplasia and in particular restenosis, smooth muscle cell inhibitors, antibiotics, growth factors, growth factor inhibitors, cell adhesion inhibitors, cell adhesion promoters and drugs that may enhance the formation of healthy neointimal tissue, including endothelial cell regeneration, etc. These agents may be encorporated in one or more coatings. Additionaly, the copper antagoinst and therapeutic agent may be combined together within the same layer or may be in separate layers.

Anti-thrombogenic agents, may include, for example, heparin, warfarin, hirudin and its analogs, aspirin, indomethacin, dipyridamole, prostacyclin, prostaglandin E, sulfinpyrazone, abciximab, eptifabatide, phenothiazines (such as chlorpromazine or trifluperazine) RGD (arginine-glycine-aspartic acid) peptide or RGD peptide mimetics, agents that block platelet glycoprotein Hb-IIIa receptors (such as C-7E3), ticlopidine or the thienopyridine known as clopidogrel.

Statins may include, for example, simvastatin, atorvastatin, lovastatin, pravastatin, and fluvastatin. α-adrenergic receptor antagonists may include, for example, prazosin, terazosin, doxazosin, ketanserin, indoramin, urapidil, clonideine, guanabenz, guanfacine, guanadrel, reserpine, and metyrosine.

^-selective adrenergic antagonist may include, for example, metoprolol, atenolol, esmolol, acebutolol, bopindolol, carteolol, oxprenolol, penbutolol, medroxalol, bucindolol, levobunolol, metipranolol, bisoprolol, nebivolol, betaxolol, celiprolol, sotalol, propafenone, propranolol, timolol maleate, and nadolol.

Vasodilators may include, for example, hydralazine, minoxidil, sodium nitroprusside, diazoxide, bosentan, eporprostenol, treprostinil, and iloprost.

(including, for example, salicylic acid derivatives i.e. aspirin; para-aminophenol derivatives e.g acetaminophen; indole and indene acetic acids, e.g. indomethacin, sulindac, and etodalac; heteroaryl acetic acids e.g. tolmetin, diclofenac, and ketorolac; arylpropionic acids e.g ibuprofen and derivatives; anthranilic acids e.g mefenamic acid, and meclofenamic acid; enolic acids e.g piroxicam, tenoxicam, phenylbutazone, and oxyphenthatrazone); and nabumetone.

Imunnosuppresant agents may include, for example, sirolimus, cyclosporine, tacrolimus, azathioprine, and mycophenolate mofetil.

Anti-proliferative/antimitotic agents may include, for example, vinca alkaloids (e.g vinblastine, vincristine, and vinorelbine), paclitaxel, epidipodophyllotoxins (e.g etoposide, teniposide), antibiotics (dactinomycin

(actinomycin D) daunorubicin, doxorubicin and idarabicin), anthracyclines, mitoxantrone, bleomycins, plicamycin (mithramycin) and mitomycin. Antiplatelet agents may include, for example, acetylsalicylic acid, dipyridamole, clopidogrel, ticlopidine, abciximab, eptifbatide, tirofiban, reversable COX-I inhibitors, BPIIIb/IIIa blockers, TP antagonists, and P2Y12 antagonists. Appropriate dose levels for such therapeutic agents are known to those in the pharmaceutical arts. Such therapeutic agents may be prepared and formulated for inclusion in the medical devices of the present invention using techinques known to those in the art.

A better understanding of the invention will be gained by reference to the following experimental section. The following experiments are illustrative of the present invention and are not intended to limit the invention in any way.

EXAMPLES

EXAMPLE 1

Determination of Effects from Use of a Copper Antagonist-Treated Device in an Animal Model

The following experiment is directed to coating artificial surfaces with a copper antagonist for amelioration of tissue damage and/or enhanced tissue repair and their testing in an animal model.

Materials: Sodium bicarbonate, sodium chloride, sodium phosphate, sodium nitrite, potassium phosphate-monobasic, 40% formaldehyde solution and sucrose are available from Fischer Scientific, Fairlawn, NJ. Sephadex G25 is available from Pharmacia, Piscataway, NJ. Monoclonal mouse anti-proliferating cell nuclear antigen is available from Dako A/S, Denmark. All other chemicals are available from Sigma Chemical Co., St. Louis, Mo. Tris-buffered saline consisted of 10 mM tris[hydroxymethyl]aminoethane, pH 7.4, and 150 mM NaCl. Phosphate-buffered saline contained 10 mM sodium phosphate and 150 mM NaCl, pH 7.4.

S-triethylenetetramine-species: S-triethylenetetramine-BSA is synthesized as follows: Fatty acid-free bovine serum albumin (200 mg/ml) is exposed to a 1.4 molar-fold excess of S-triethylenetetramine in 0.5 M HCl for 30 minutes at room temperature and neutralized with an equal volume of TBS and 0.5 M NaOH.

Thiolated bovine serum albumin (pS-BSA) is prepared after Benesch and Benesch (Benesch R & Benesch RE, "Preparation and properties of hemoglobin modified with derivatives of pyridoxal". Methods Enzymol., 76:147-159 (1981)). Briefly, essential fatty acid-free bovine serum albumin (50 mg/ml) is dissolved in water with N-acetyl-homocysteine thiolactone (35 mM) and 0.05% polyethylenesorbitan monolaurate. Equimolar Silver nitrate is slowly added at room temperature over 90 minutes at pH 8.5. Excess thiourea (70 mM) is added and the pH lowered to 2.5. Excess silver nitrate is removed by Dowex 50 chromatography with the mobile phase consisting of IM thiourea, pH 2.5, and excess thiourea is removed by Sephadex G-25 chromatography. The pS-BSA is prepared within two days of subsequent derivitization and stored at 4⁰C. Derivitization of pS-BSA is accomplished with 1.4 fold molar excess S- triethylenetetramine in 0.5 N HCl for 30 minutes at room temperature. The solution is adjusted to pH 4.0 with 0.5 N NACH after derivitization. Protein content is determined using the method of Lowry and colleagues (Marcus Salier, FASEBJ., 7:516-522, 1993).

Animal Preparation: All animal preparations are performed within institutional guidelines of the host institution. New Zealand white rabbits (3.5- 4.2 kg) of either sex are premedicated with 5 mg/kg intramuscular (IM) xylazine hydrochlolide (Miles Pharmaceuticals, Shawnee Mission, Kans.), and 0.1 mg/kg subcutaneous (SC) atropine sulfate (Lyphomed, Deerfield, 111.) fifteen minutes prior to the induction of anesthesia. Anaesthesia is induced with 40 mg/kg IM ketamine hydrochloride (Fort Dodge Laboratories, Fort Dodge, Iowa) and 5 mg/kg IM acepromazine maleate (Aveco Company, Inc., Fort Dodge, Iowa). Additional doses of ketamine hydrochloride are administered as necessary to maintain anesthesia. For survival studies, 100,000 U penicillin G (Apothecon of Bristol-Myers Squibb, Princeton, NJ.), is administered IM perioperatively. The skin over the femoral arteries is infiltrated with 1% lidocaine (Astra Pharmaceuticals, Inc., Westborough, Mass.), and the common femoral arteries are exposed from the inguinal ligament to the superficial femoral artery. Arteries are cleared of connective tissue, side branches are ligated, and the superficial femoral artery is suspended with silk ties. A 1.5-to-2.0 cm length of femoral artery is isolated from the circulation proximally and distally with neurosurgical microaneurysm clips. The superficial femoral artery is cannulated with a S- triethylenetetramine-BSA coated 2F Fogarty balloon catheter (American Edwards Laboratories, Santa Ana, Calif), that is passed into the isolated segment of femoral artery. The balloon is inflated with sufficient air to generate slight resistance and withdrawn three times. The contralateral femoral artery is prepared identically as an appropriate control, i.e., using a balloon catheter coated with underivatized BSA. Following removal of the balloon catheter, the superficial femoral artery is ligated and flow re-established. The area of balloon injury is marked by surgical staples in the adjacent muscle fascia. The incision is closed with subcuticular absorbable suture and the animals allowed to recover. In some experiments, a distant control vessel, the right carotid artery, is isolated and harvested without any other manipulation.

Tissue processing and analysis: On the 14th postoperative day, animals are euthanized with 120 mg/kg intravenous sodium pentobarbital (Anpro Pharmaceuticals, Arcadia, Calif), and the abdominal aorta and inferior vena cava interrupted by silk ties. A 7F plastic cannula is inserted into the abdominal aorta and the vessels perfused clear with saline followed by fixation at 100 mm Hg pressure with 10% buffered formalin. The vessels are stored in 10% buffered formalin and the samples paraffin-embedded and microtome-sectioned. Six sections are made along the length of each injured segment of vessel and stained with Verhoeff s stain for elastic tissue. The areas within the lumen, internal elastic membrane, and external elastic membrane are measured by a blinded observer using computerized digital planimetry (Zeiss, West Germany). The areas within the lumen, internal elastic membrane and external elastic membrane are analyzed. Sections with obstructive thrombus impairing analysis are discarded. In a separate set of animals, vessels are perfosion-fϊxed with 10% buffered formalin seven days after injury and processed for analysis of proliferating cells within 12 hours. Sections are stained for proliferating cell nuclear antigen (PCNA) and adjacent sections are stained with hematoxylin and eosin. Five representative sections from each segment are examined. Total nuclei are counted from the hematoxylin and eosin slides and percent PCNA positive cells are defined as the number of PCNA-positive nuclei divided by the total number of nuclei multiplied by 100.

Statistics: Treatments are administered in a paired fashion with one femoral artery cannulated with an S-triethylenetetramine-BSA coated balloon catheter while the other femoral artery is cannulated with an underivatized BSA coated balloon catheter. Data is tested for normality using appropriate statistical methods, for example, the Kolmogorov-Smirnov algorithm and for equal variance with the Levene Median test. Normally distributed variables are compared using, for example, the paired t-test and non-normally distributed variables using, for example, the Wilcoxon sign-ranks test or the Mann- Whitney rank-sum test. Non-paired data are compared using, for example, an independent t-test. Statistical significance is accepted if the null hypothesis is rejected with P<0.05. S-triethylenetetramine-BSA effect on platelet binding to injured vessel:

Platelet adhesion to the injured arterial surface has been reported to be important in the proliferative response to injury, and the effects of S-triethylenetetramine- BSA on platelet deposition after balloon injury are investigated. Platelet deposition is assessed at the site of cannulation with a S-triethylenetetramine- BSA coated balloon catheter, and compared to that at the site of cannulation with an underivatized BSA coated balloon catheter. Decreased platelet deposition at the site of cannulation with S-triethylenetetramine-BSA coated balloon catheter is indicatitve of ameliorated tissue damage and/or enhanced tissue repair.

S-triethylenetetramine-BSA effects on neointimal proliferation: Neointimal proliferation after local delivery of S-triethylenetetramine-BSA and appropriate controls are evaluated by comparing absolute neointimal area and neointima/media ratios. The absolute neointimal area and neointima/media ratio at the site of cannulation with an S-triethylenetetramine-BSA coated balloon catheter is assessed and compared to a neointimal area and neointima/media ratio at the site of cannulation with an underivatized BSA coated balloon catheter. A decrease in neointimal area or reduction in the neointima/media ratio is indicative of an inhibition of neointimal proliferation and of amelioration of tissue damage and/or enhanced tissue repair.

S-triethylenetetramine-BSA effects on cellular proliferation: Mouse monoclonal antibody staining against PCNA is used to assay the degree of Sl- phase activity at 7 days after injury. At this time, the percent of proliferating cells is assessed in vessels cannulated with a S-triethylenetetramine-BSA coated balloon catheter and in vessels cannulated with an underivatized BSA coated balloon catheter. Histological assessment determines the cellular populations undergoing proliferation. These experiments are directed to the effect on neointimal proliferation and amelioration of tissue damage and/or enhanced tissue repair by localised delivery of a copper antagonist by, for example, the coating of a balloon catheter with S- triethy lenetetramine-B SA .

The endothelium is reported to be essential for vascular integrity, control of thrombosis, (Clowes et al, Lab. Invest. 49:327-333, 1983); (Rees et al., Proc. Natl. Acad. Sci. USA. 86:3375-3378, 1989) and the regulation of intimal growth (Kubes et al., Proc. Natl. Acad. Sci. USA, 88:4651-4655, 1991), and has been proposed to be important in the local control of vascular smooth muscle growth. Balloon angioplasty reportedly removes the endothelium from arterial smooth muscle, and these endothelial functions can often be lost during the procedure. In particular, removal of the endothelium and damage to the smooth muscle cells have been reported to be associated with intimal proliferation (McNamara et al., Biochem. Biophys. Res. Commun., 193:291-296, 1993). The mechanism for this response is complex and reportedly involves platelet deposition and activation, cytokine elaboration, smooth muscle cell migration and proliferation, and extracellular matrix production. It has been reported that after balloon injury, the endothelium regenerates rapidly but is often dysfunctional, (Saville, Analyst 83:670-672, 1958).

A limitation of neointimal proliferation after a single, local administration of a copper antagonist is indicative of amelioration of tissue damage and/or enhanced tissue repair. Antiplatelet activity may explain such findings. Inhibition of platelet binding has been said to result in many effects that are likely to reduce the proliferative response after injury. For example, platelet adhesion and aggregation is said to be associated with the release of PDGF, basic fibroblast growth factor, epidermal growth factor, and transforming growth factor-β, potent stimuli for smooth muscle cell proliferation and matrix production.

There may additionally or alternatively be a direct effect on vascular smooth muscle gene expression, migration, proliferation or synthesis of extracellular matrix. Such an effect may be in addition to an amelioration of tissue damage an/or enhancement in tissue repair by, for example, restoration of normal tissue stem cell responses.

It has been reported that the mechanical removal of the endothelium abolishes the vasodilator responses to endothelium-dependent vasoactive stimuli, while leaving the vasoconstrictor effects of agonists to smooth muscle unopposed (Furchgott Zawadzki, Nature (Lond.)., 288:373-376 (1980). This process reportedly occurs with balloon angioplasty especially at sites where platelet thrombus is noted (Uchida et al., Am. Heart. J., 117:769-776 (1989); Steele et al., Circ. Res., 57:105-112 (1985). The strategy of administration of a copper antagonist as therapy for, for example, acute thrombotic phenomena and restenosis following angioplasty is supported by results indicating an amelioration of tissue damage and/or enhancement in tissue repair. EXAMPLE 2

Preparation and Use of a Copper Antagonist Coated Medical Device for Ameliorating Tissue Damage and/or Enhancing Tissue Repair The following experiment is directed to application of copper antagonists to coated artificial surfaces, such as, for example, synthetic vascular graft material, or stents, for use in ameliorating tissue damage and/or enhancing tissue repair.

First, dacron grafts and cardiac catheters are coated with a functionalized copper antagonist, such as for example, S-triethylenetetramine-bovine serum albumin (BSA). In three separate experiments, an identical pair of 6 mm (internal diameter) knitted dacron grafts, 5 cm in length, are prepared for surgical placement in the transected carotid arteries of six anesthetized dogs. For each experiment, three control dogs receive grafts coated in underivatized BSA, while for each of the three trial dogs, one graft is soaked in 5% BSA and the other graft is soaked in 5% BSA combined with 0.5 mM S-triethylenetetramine producing, S- triethylenetetramine-BSA, an example of a functionalized copper antagonist protein, for one hour prior to insertion, and then rinsed in saline. The grafts are sutured in place with a continuous 6-0 proline suture.

Following graft insertion, the dogs are observed for two months. During this two month observation, copper excretion in urine is monitored. Administration of triethylenetetramine has been reported to increase copper excretion in the urine.

Histological examination of the site of insertion is performed at the end of the two month period. Amelioration of tissue damage or enhanced tissue repair is evidenced by, for example, the appearance of normal vascular epithelial cells at the site of graft insertion. Tissue repair at the site of S-triethylenetetramine- BSA-coated graft insertion is compared to that at the site of underivatized, BSA- coated graft insertion.

Evidence of amelioration of tissue damage and/or enhanced tissue repair at the site of S -triethylenetetramine-BSA coated graft insertion compared to that at the site of underivatized BSA-coated graft insertion is shown by amelioration of tissue damage and/or enhanced tissue repair at the site of insertion of the synthetic grafts treated with S-triethylenetetramine-BSA during exposure of the graft to circulating blood over a period of two months.

EXAMPLE 3

Application of Copper Agonists to Damaged Vascular Surfaces to Ameliorate Tissue Damage and/or Enhance Tissue Repair

The following experiment is directed to application of copper antagonists, such as the functionalized compounds of Formula I or II or, for example, S- triethylenetetramine-BSA, to damaged vascular surfaces, for example, arterial surfaces, to ameliorate tissue damage and/or enhance tissue repair.

In five anesthetized dogs, both carotid arteries are exposed. Two 3 FR USCl catheters are prepared for arterial implantation. One catheter is soaked in a 5% BSA solution for 12 hours, while the other is soaked in a 5% BSA solution which also contains 1 mg/ml of S-triethylenetetramine. One each of the two coated catheters is placed randomly in the right or left carotid artery of the dog through a small incision sealed with a 6-0 proline suture. The catheters are advanced for 5 cm into the arterial lumen. Following catheter insertion, the dogs are observed for two weeks. During this two week observation, copper excretion in urine is monitored.

Histological examination of the site of insertion is performed at the end of the two week period. Lessened tissue damage, and/or tissue repair, is evidenced by, for example, the appearance of normal vascular epithelial cells at the site of catheter insertion. Tissue at the site of S-triethylenetetramine-BSA coated catheter insertion is compared to that at the site of underivatized BSA-coated catheter insertion.

Evidence of enhanced tissue repair and/or amelioration of tissue damage at the site of S-triethylenetetramine-BSA coated catheter insertion compared to that at the site of underivatized BSA-coated catheter insertion is shown by enhanced tissue repair and/or amelioration of tissue damage at the site of insertion of the catheter treated with S-triethylenetetramine-BSA during exposure of the catheter to circulating blood over a period of two weeks.

EXAMPLE 4

Determination of Tissue Damage Using a Copper Antagonist Treated Medical Device in a Pig Model

The following experiment is directed to determining the tissue damage using a treated medical device in a pig model.

Pigs are subjected to coronary balloon-injury using standard methods. Prior to balloon injury, an angiogram is performed. Thereafter, in trial pigs, an S- triethylenetetramine-BSA coated balloon catheter, for example, is used for balloon injury, whereas in control pigs, an underivatized BSA coated balloon catheter is used. The balloon of the catheter is inflated for 15 min, then deflated and the catheter is removed. Another angiogram is performed 30 minutes after injury to determine the degree of spasm. Coronary catheters are placed in the coronary ostea, radiocontrast is infused into the coronary arteries and measurements are made of the degree of so-called "recoil spasm" that exists at the point of angioplasty. The degree of spasm or recoil is defined quantitatively, again using the computer-driven quantitative coronary angiography algorithm that compares the segment at the site of balloon injury with a proximal segment that is uninjured as a reference standard. All catheters are then removed and incision sites repaired. The animals are awakened and maintained with normal chow diets over the next four weeks. At the end of that period of time, the animals are again sedated, undergo coronary angiography to determine coronary stenoses at the site of angioplasty. Catheters are placed in the coronary ostea and radiocontrast fluid is infused. The angiograms are recorded and subsequently processed by a computer-driven quantitative coronary angiography algorithm to determine lumen diameter. The degree of stenosis represents the percentage reduction in the lumen diameter compared with a reference segment proximal to the area of stenosis using standard methods. The animals are euthanized by an overdose of pentobarbital. Their coronary arteries are perfusion fixed with formalin at 100 mm Hg of perfusion pressure, harvested and sectioned for quantitative morphometric assessment of the lumen diameter, the neointimal dimension and cross-section, as well as the neointimal area. The arteries are stained with hematoxylin and eosin. The neointima to lumen diameter ratio is determined and is compared between trial and control animals.

EXAMPLE 5

Determination of Effect on Tissue Damage and/or Tissue Repair Using a Copper Antagonist Coated Medical Device This experiment is directed to coating a Palmaz-Schatz stent with a copper antagonist to reduce the degree and severity of neointimal hyperplasia leading to restenosis. Palmaz-Schatz stents are dip-coated in 800-1000μM S- triethylenetetramine-BSA, for example, or underivatized BSA three times for 10 minutes followed by 10 minutes of air drying time. One S-triethylenetetramine- BSA coated and one underivatized BSA-coated stent is placed under sterile conditions in the carotid arteries of 10 pigs, one in each carotid artery. They are followed for 28 days and then the carotid arteries are removed. They are examined histologically for the degree of neointimal hyperplasia, and tissue damage/repair. Tissue repair is evidenced by, for example, the appearance of normal vascular epithelial cells at the site of insertion of the stent. Tissue damage/repair at the site of S-triethylenetetramine-BSA coated stent insertion is compared to that at the site of underivatized BSA-coated stent insertion.

Evidence of amelioration of tissue damage and/or enhanced tissue repair at the site of S-triethylenetetramine-BSA coated stent insertion compared to that at the site of underivatized BSA-coated stent insertion shows that during exposure of the stent to circulating blood, there is amelioration of tissue damage and/or enhanced tissue repair at the site of insertion of the stent coated with copper antagonist, and that delivery of a copper antagonist is able to ameliorate tissue damage enhance tissue repair. Φ** Φ All patents, publications, scientific articles, web sites, and other documents and materials referenced or mentioned herein are indicative of the level of skill of those skilled in the art to which the invention pertains, and each such referenced document and material is hereby incorporated by reference to the same extent as if it had been incorporated by reference in its entirety individually or set forth herein in its entirety. Applicants reserve the right to physically incorporate into this specification any and all materials and information from any such patents, publications, scientific articles, web sites, electronically available information, and other referenced materials or documents. The written description portion of this patent includes all claims.

Furthermore, all claims, including all original claims as well as all claims from any and all priority documents, are hereby incorporated by reference in their entirety into the written description portion of the specification, and Applicants reserve the right to physically incorporate into the written description or any other portion of the application, any and all such claims. Thus, for example, under no circumstances may the patent be interpreted as allegedly not providing a written description for a claim on the assertion that the precise wording of the claim is not set forth in haec verba in written description portion of the patent.

The claims will be interpreted according to law. However, and notwithstanding the alleged or perceived ease or difficulty of interpreting any claim or portion thereof, under no circumstances may any adjustment or amendment of a claim or any portion thereof during prosecution of the application or applications leading to this patent be interpreted as having forfeited any right to any and all equivalents thereof that do not form a part of the prior art.

All of the features disclosed in this specification may be combined in any combination. Thus, unless expressly stated otherwise, each feature disclosed is only an example of a generic series of equivalent or similar features.

It is to be understood that while the invention has been described in conjunction with the detailed description thereof, the foregoing description is intended to illustrate and not limit the scope of the invention, which is defined by the scope of the appended claims. Thus, from the foregoing, it will be appreciated that, although specific embodiments of the invention have been described herein for the purpose of illustration, various modifications may be made without deviating from the spirit and scope of the invention. Other aspects, advantages, and modifications are within the scope of the following claims and the present invention is not limited except as by the appended claims.

The specific methods and compositions described herein are representative of preferred embodiments and are exemplary and not intended as limitations on the scope of the invention. Other objects, aspects, and embodiments will occur to those skilled in the art upon consideration of this specification, and are encompassed within the spirit of the invention as defined by the scope of the claims. It will be readily apparent to one skilled in the art that varying substitutions and modifications may be made to the invention disclosed herein without departing from the scope and spirit of the invention. The invention illustratively described herein suitably may be practiced in the absence of any element or elements, or limitation or limitations, not specifically disclosed herein as essential. Thus, for example, in each instance herein, in embodiments or examples of the present invention, the terms "comprising", "including", "containing", etc. are to be read expansively and without limitation. In particular, the phrase "for example" shall be interpreted to mean "for example, and including but not limited to." The methods and processes illustratively described herein suitably may be practiced in differing orders of steps, and that they are not necessarily restricted to the orders of steps indicated herein or in the claims. The terms and expressions that have been employed are used as terms of description and not of limitation, and there is no intent in the use of such terms and expressions to exclude any equivalent of the features shown and described or portions thereof, but it is recognized that various modifications are possible within the scope of the invention as claimed. Thus, it will be understood that although the present invention has been specifically disclosed by various embodiments and/or preferred embodiments and optional features, any and all modifications and variations of the concepts herein disclosed that may be resorted to by those skilled in the art are considered to be within the scope of this invention as defined by the appended claims.

The invention has been described broadly and generically herein. Each of the narrower species and subgeneric groupings falling within the generic disclosure also form part of the invention. This includes the generic description of the invention with a proviso or negative limitation removing any subject matter from the genus, regardless of whether or not the excised material is specifically recited herein. It is also to be understood that as used herein and in the appended claims, the singular forms "a", "an", and "the" include plural reference unless the context clearly dictates otherwise, the term "X and/or Y" means "X" or "Y" or both "X" and "Y", and the letter "s" following a noun designates both the plural and singular forms of that noun. In addition, where features or aspects of the invention are described in terms of Markush groups, those skilled in the art will recognize that the invention is also thereby described in terms of any individual member or subgroup of members of the Markush group.

Other embodiments are within the following claims. Under no circumstances may the patent be interpreted to be limited to the specific examples or embodiments or methods specifically and/or expressly disclosed herein. Under no circumstances may the patent be interpreted to be limited by any statement made by any Examiner or any other official or employee of the Patent and Trademark Office unless such statement is specifically and without qualification or reservation expressly adopted in a responsive writing by Applicants.

Claims

CLAIMS:

1. An implantable medical device comprising a copper antagonist releasable upon insertion of the medical device to or within a subject.

2. The medical device of claim 1, wherein the copper antagonist chelates copper.

3. The medical device of claim 1 or claim 2, wherein the copper antagonist binds copper (II).

4. The medical device according to any one of claims 1 to 3, wherein the copper antagonist binds copper (II) by chelation.

5. The medical device according to any one of claims 1 to 4, wherein the copper antagonist is triethylenetetramine or a pharmaceutically acceptable salt thereof.

6. The medical device according to any one of claims 1 to 5, wherein the copper antagonist is selected from the group consisting of a triethylenetetramine hydrochloride salt, a triethylenetetramine succinate salt, a triethylenetetramine fumarate salt, and a triethylenetetramine maleate salt.

7. The medical device according to any one of claims 1 to 6, wherein the copper antagonist is a triethylenetetramine disuccinate salt.

8. The medical device of claim 1, wherein the copper antagonist is a compound of Formula I.

9. The medical device of claim 1, wherein the copper antagonist is a compound of Formula II.

10. The medical device of claim 1, wherein the copper antagonist is selected from the group consisting of:

SH-CH₂-CH₂-NH-CH₂-CH₂-NH-CH₂-CH₂-NH₂, SH-CH₂-CH₂-S-CH₂-CH₂-NH-CH₂-CH₂-NH₂₅

NH₂-CH₂-CH₂-NH-CH₂-CH₂-S-CH₂-CH₂-SH, NH₂-CH₂-CH₂-S-CH₂-CH₂-S-CH₂-CH₂-SH, SH-CH₂-CH₂-S-CH₂-CH₂-S-CH₂-CH₂-SH, NH₂-CH₂-CH₂-NH-CH₂-CH₂-CH₂-NH-CH₂-CH₂-NH₂,

SH-CH₂-CH₂-NH-CH₂-CH₂-CH₂-NH-CH₂-CH₂-NH₂, SH-CH₂-CH₂-S-CH₂-CH₂-CH₂-NH-CH₂-CH₂-NH₂, NH₂-CH₂-CH₂-NH-CH₂-CH₂-CH₂-S-CH₂-CH₂-SH, NH₂-CH₂-CH₂-S-CH₂-CH₂-CH₂-S-CH₂-CH₂-SH, and SH-CH₂-CH₂-S-CH₂-CH₂-CH₂-S-CH₂-CH₂-SH.

11. The medical device according to any one of claims 1 to 10, wherein the surface of the medical device comprises a copper antagonist.

12. The medical device of claim 11, wherein the surface of the medical device contacts a target tissue within the subject upon use.

13. The medical device of claim 12, wherein the target tissue is heart tissue or vascular tissue.

14. The medical device according to any one of claims 1 to 13, wherein the copper antagonist is present at a weight percentage of about 0.0001% to about 30%.

15. The medical device according to any one of claims 1 to 14, wherein the release rate of the copper antagonist is controlled.

16. The medical device according to any one of claims 1 to 15, wherein the medical device provides for surface contact release of the copper antagonist.

17. The medical device according to any one of claims \ to 15, wherein the medical device provides for the sustained release of the copper antagonist.

18. The medical device according to any one of claims 1 to 15, wherein the medical device provides for the slow release of the copper antagonist.

19. The medical device according to any one of claims 1 to 18, wherein the device comprises a coating containing a copper antagonist.

20. The medical device of claim 19, wherein the coating comprises a polymer.

21. The medical device of claim 19 wherein the coating comprises a plurality of layers of a polymer/copper antagonist mixture.

22. The medical device according to any one of claims 1 to 21, wherein the medical device is for implantation in a human.

23. The medical device according to any one of claims 1 to 21, wherein the medical device comprises a stent.

24. The medical device of claim 23 wherein the stent is a drug-eluting stent.

25. The medical device according to any one of claims 1 to 21, wherein the medical device comprises a balloon, a prosthetic heart valve, an annuloplasty ring, a pulse generator, a cardiac defibrillator, an arteriovenous shunt, an anastomosis device, a hemostatic barrier or a pacemaker.

26. The medical device according to any one of claims 1 to 21, wherein the medical device comprises an orbital implant, a lens, a lens implant, or a corneal implant.

27. The medical device according to any one of claims 1 to 21, wherein the medical device comprises an orthopedic plate, a bone pin, a bone substitute, a anchor, a joint, a screw, or a vertebral disk.

28. The medical device according to any one of claims 1 to 21, wherein the medical device comprises a graft, a shunt, a vascular implant, a tissue scaffold, an intraluminal device or a vascular support.

29. The medical device according to any one of any of claims 23 to 28, wherein the medical device is for implantation in a human.

30. The medical device according to any one of claims 1 to 21, wherein the medical device comprises at least one channel formed in an outer surface thereof, and wherein the copper antagonist is included on and/or within at least one channel.

31. The medical device according to any one of claims 1 to 21, wherein at least a portion of the medical device is formed, in whole or in part, of a substance that includes the copper antagonist.

32. The medical device according to any one of claims 1 to 31, wherein the device further comprises one or more therapeutic agents.

33. The medical device according to any one of claims 30 to 32, wherein the medical device is for implantation in a human.

34. A method of preventing and/or treating damage associated with the use or implantation of a medical device in a subject comprising introducing into the subject a medical device of which at least a portion comprises a copper antagonist, wherein the damage is prevented, amerliorated and/or delayed.

35. The method of claim 34 wherein the subject is a mammal.

36. The method of claim 35 wherein the mammal is a human.

37. The method of claim 35 wherein the mammal is selected from the group consisting of domestic and pet animals, sports animals, farm animals, and zoo animals.

38. The method of claim 35 wherein the mammal is a horse, a dog, or a cat.

39. The method according to any one of claims 34 to 38, wherein the surface of said medical device contacts a site of injury or potential injury.

40. A method of preventing and/or treating damage associated with the use or implantation of a medical device in a subject comprising use or implantation of a medical device which comprises a copper antagonist that is releasable at its point of contact, wherein the damage is prevented, amerliorated and/or delayed.