CA2160693C - Oral drug delivery compositions and methods - Google Patents

Oral drug delivery compositions and methods Download PDF

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Publication number
CA2160693C
CA2160693C CA2160693A CA2160693A CA2160693C CA 2160693 C CA2160693 C CA 2160693C CA 2160693 A CA2160693 A CA 2160693A CA 2160693 A CA2160693 A CA 2160693A CA 2160693 C CA2160693 C CA 2160693C
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Prior art keywords
alkyl
alkenyl
phenyl
naphthyl
acid
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CA2160693A
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French (fr)
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CA2160693A1 (en
Inventor
Sam J. Milstein
Evgueni Barantsevitch
Donald J. Sarubbi
Andrea Leone-Bay
Duncan R. Paton
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Emisphere Technologies Inc
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Emisphere Technologies Inc
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Priority claimed from US08/051,019 external-priority patent/US5451410A/en
Priority claimed from US08/205,511 external-priority patent/US5792451A/en
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    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00Medicinal preparations containing organic active ingredients
    • A61K31/16Amides, e.g. hydroxamic acids
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00Medicinal preparations containing organic active ingredients
    • A61K31/33Heterocyclic compounds
    • A61K31/335Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
    • A61K31/35Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom
    • A61K31/352Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom condensed with carbocyclic rings, e.g. methantheline 
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00Medicinal preparations containing organic active ingredients
    • A61K31/70Carbohydrates; Sugars; Derivatives thereof
    • A61K31/715Polysaccharides, i.e. having more than five saccharide radicals attached to each other by glycosidic linkages; Derivatives thereof, e.g. ethers, esters
    • A61K31/726Glycosaminoglycans, i.e. mucopolysaccharides
    • A61K31/727Heparin; Heparan
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00Medicinal preparations containing peptides
    • A61K38/16Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/17Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • A61K38/19Cytokines; Lymphokines; Interferons
    • A61K38/21Interferons [IFN]
    • A61K38/212IFN-alpha
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00Medicinal preparations containing peptides
    • A61K38/16Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/17Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • A61K38/22Hormones
    • A61K38/23Calcitonins
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00Medicinal preparations containing peptides
    • A61K38/16Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/17Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • A61K38/22Hormones
    • A61K38/25Growth hormone-releasing factor [GH-RF] (Somatoliberin)
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00Medicinal preparations containing peptides
    • A61K38/16Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/17Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • A61K38/22Hormones
    • A61K38/27Growth hormone [GH] (Somatotropin)
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00Medicinal preparations characterised by special physical form
    • A61K9/14Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles
    • A61K9/16Agglomerates; Granulates; Microbeadlets ; Microspheres; Pellets; Solid products obtained by spray drying, spray freeze drying, spray congealing,(multiple) emulsion solvent evaporation or extraction
    • A61K9/1605Excipients; Inactive ingredients
    • A61K9/1617Organic compounds, e.g. phospholipids, fats
    • CCHEMISTRY; METALLURGY
    • C07ORGANIC CHEMISTRY
    • C07CACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C229/00Compounds containing amino and carboxyl groups bound to the same carbon skeleton
    • C07C229/40Compounds containing amino and carboxyl groups bound to the same carbon skeleton having amino groups bound to carbon atoms of at least one six-membered aromatic ring and carboxyl groups bound to acyclic carbon atoms of the same carbon skeleton
    • C07C229/42Compounds containing amino and carboxyl groups bound to the same carbon skeleton having amino groups bound to carbon atoms of at least one six-membered aromatic ring and carboxyl groups bound to acyclic carbon atoms of the same carbon skeleton with carboxyl groups linked to the six-membered aromatic ring, or to the condensed ring system containing that ring, by saturated carbon chains
    • CCHEMISTRY; METALLURGY
    • C07ORGANIC CHEMISTRY
    • C07CACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C233/00Carboxylic acid amides
    • C07C233/01Carboxylic acid amides having carbon atoms of carboxamide groups bound to hydrogen atoms or to acyclic carbon atoms
    • C07C233/45Carboxylic acid amides having carbon atoms of carboxamide groups bound to hydrogen atoms or to acyclic carbon atoms having the nitrogen atom of at least one of the carboxamide groups bound to a carbon atom of a hydrocarbon radical substituted by carboxyl groups
    • C07C233/53Carboxylic acid amides having carbon atoms of carboxamide groups bound to hydrogen atoms or to acyclic carbon atoms having the nitrogen atom of at least one of the carboxamide groups bound to a carbon atom of a hydrocarbon radical substituted by carboxyl groups with the substituted hydrocarbon radical bound to the nitrogen atom of the carboxamide group by a carbon atom of a six-membered aromatic ring
    • C07C233/55Carboxylic acid amides having carbon atoms of carboxamide groups bound to hydrogen atoms or to acyclic carbon atoms having the nitrogen atom of at least one of the carboxamide groups bound to a carbon atom of a hydrocarbon radical substituted by carboxyl groups with the substituted hydrocarbon radical bound to the nitrogen atom of the carboxamide group by a carbon atom of a six-membered aromatic ring having the carbon atom of the carboxamide group bound to a carbon atom of an unsaturated carbon skeleton
    • CCHEMISTRY; METALLURGY
    • C07ORGANIC CHEMISTRY
    • C07CACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C233/00Carboxylic acid amides
    • C07C233/57Carboxylic acid amides having carbon atoms of carboxamide groups bound to carbon atoms of rings other than six-membered aromatic rings
    • C07C233/63Carboxylic acid amides having carbon atoms of carboxamide groups bound to carbon atoms of rings other than six-membered aromatic rings having the nitrogen atom of at least one of the carboxamide groups bound to a carbon atom of a hydrocarbon radical substituted by carboxyl groups
    • CCHEMISTRY; METALLURGY
    • C07ORGANIC CHEMISTRY
    • C07CACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C233/00Carboxylic acid amides
    • C07C233/64Carboxylic acid amides having carbon atoms of carboxamide groups bound to carbon atoms of six-membered aromatic rings
    • C07C233/81Carboxylic acid amides having carbon atoms of carboxamide groups bound to carbon atoms of six-membered aromatic rings having the nitrogen atom of at least one of the carboxamide groups bound to a carbon atom of a hydrocarbon radical substituted by carboxyl groups
    • C07C233/82Carboxylic acid amides having carbon atoms of carboxamide groups bound to carbon atoms of six-membered aromatic rings having the nitrogen atom of at least one of the carboxamide groups bound to a carbon atom of a hydrocarbon radical substituted by carboxyl groups with the substituted hydrocarbon radical bound to the nitrogen atom of the carboxamide group by an acyclic carbon atom
    • C07C233/87Carboxylic acid amides having carbon atoms of carboxamide groups bound to carbon atoms of six-membered aromatic rings having the nitrogen atom of at least one of the carboxamide groups bound to a carbon atom of a hydrocarbon radical substituted by carboxyl groups with the substituted hydrocarbon radical bound to the nitrogen atom of the carboxamide group by an acyclic carbon atom of a carbon skeleton containing six-membered aromatic rings
    • CCHEMISTRY; METALLURGY
    • C07ORGANIC CHEMISTRY
    • C07CACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C235/00Carboxylic acid amides, the carbon skeleton of the acid part being further substituted by oxygen atoms
    • C07C235/02Carboxylic acid amides, the carbon skeleton of the acid part being further substituted by oxygen atoms having carbon atoms of carboxamide groups bound to acyclic carbon atoms and singly-bound oxygen atoms bound to the same carbon skeleton
    • C07C235/32Carboxylic acid amides, the carbon skeleton of the acid part being further substituted by oxygen atoms having carbon atoms of carboxamide groups bound to acyclic carbon atoms and singly-bound oxygen atoms bound to the same carbon skeleton the carbon skeleton containing six-membered aromatic rings
    • C07C235/38Carboxylic acid amides, the carbon skeleton of the acid part being further substituted by oxygen atoms having carbon atoms of carboxamide groups bound to acyclic carbon atoms and singly-bound oxygen atoms bound to the same carbon skeleton the carbon skeleton containing six-membered aromatic rings having the nitrogen atom of at least one of the carboxamide groups bound to a carbon atom of a six-membered aromatic ring
    • CCHEMISTRY; METALLURGY
    • C07ORGANIC CHEMISTRY
    • C07CACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C235/00Carboxylic acid amides, the carbon skeleton of the acid part being further substituted by oxygen atoms
    • C07C235/42Carboxylic acid amides, the carbon skeleton of the acid part being further substituted by oxygen atoms having carbon atoms of carboxamide groups bound to carbon atoms of six-membered aromatic rings and singly-bound oxygen atoms bound to the same carbon skeleton
    • C07C235/44Carboxylic acid amides, the carbon skeleton of the acid part being further substituted by oxygen atoms having carbon atoms of carboxamide groups bound to carbon atoms of six-membered aromatic rings and singly-bound oxygen atoms bound to the same carbon skeleton with carbon atoms of carboxamide groups and singly-bound oxygen atoms bound to carbon atoms of the same non-condensed six-membered aromatic ring
    • C07C235/58Carboxylic acid amides, the carbon skeleton of the acid part being further substituted by oxygen atoms having carbon atoms of carboxamide groups bound to carbon atoms of six-membered aromatic rings and singly-bound oxygen atoms bound to the same carbon skeleton with carbon atoms of carboxamide groups and singly-bound oxygen atoms bound to carbon atoms of the same non-condensed six-membered aromatic ring with carbon atoms of carboxamide groups and singly-bound oxygen atoms, bound in ortho-position to carbon atoms of the same non-condensed six-membered aromatic ring
    • C07C235/64Carboxylic acid amides, the carbon skeleton of the acid part being further substituted by oxygen atoms having carbon atoms of carboxamide groups bound to carbon atoms of six-membered aromatic rings and singly-bound oxygen atoms bound to the same carbon skeleton with carbon atoms of carboxamide groups and singly-bound oxygen atoms bound to carbon atoms of the same non-condensed six-membered aromatic ring with carbon atoms of carboxamide groups and singly-bound oxygen atoms, bound in ortho-position to carbon atoms of the same non-condensed six-membered aromatic ring having the nitrogen atom of at least one of the carboxamide groups bound to a carbon atom of a six-membered aromatic ring
    • CCHEMISTRY; METALLURGY
    • C07ORGANIC CHEMISTRY
    • C07CACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C235/00Carboxylic acid amides, the carbon skeleton of the acid part being further substituted by oxygen atoms
    • C07C235/70Carboxylic acid amides, the carbon skeleton of the acid part being further substituted by oxygen atoms having carbon atoms of carboxamide groups and doubly-bound oxygen atoms bound to the same carbon skeleton
    • C07C235/84Carboxylic acid amides, the carbon skeleton of the acid part being further substituted by oxygen atoms having carbon atoms of carboxamide groups and doubly-bound oxygen atoms bound to the same carbon skeleton with the carbon atom of at least one of the carboxamide groups bound to a carbon atom of a six-membered aromatic ring
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    • C07ORGANIC CHEMISTRY
    • C07CACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C237/00Carboxylic acid amides, the carbon skeleton of the acid part being further substituted by amino groups
    • C07C237/02Carboxylic acid amides, the carbon skeleton of the acid part being further substituted by amino groups having the carbon atoms of the carboxamide groups bound to acyclic carbon atoms of the carbon skeleton
    • C07C237/22Carboxylic acid amides, the carbon skeleton of the acid part being further substituted by amino groups having the carbon atoms of the carboxamide groups bound to acyclic carbon atoms of the carbon skeleton having nitrogen atoms of amino groups bound to the carbon skeleton of the acid part, further acylated
    • CCHEMISTRY; METALLURGY
    • C07ORGANIC CHEMISTRY
    • C07CACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C2601/00Systems containing only non-condensed rings
    • C07C2601/02Systems containing only non-condensed rings with a three-membered ring
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    • C07ORGANIC CHEMISTRY
    • C07CACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C2601/00Systems containing only non-condensed rings
    • C07C2601/12Systems containing only non-condensed rings with a six-membered ring
    • C07C2601/14The ring being saturated
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    • C07C2601/00Systems containing only non-condensed rings
    • C07C2601/18Systems containing only non-condensed rings with a ring being at least seven-membered

Abstract

The present invention relates to an oral drug delivery system, and in particular to modified amino acids and modified amino acid derivatives for use as a delivery system of sensitive agents such as bioactive peptides. The modified amino acids and derivatives can form non-covalent mixtures with active biological agents and in an alternate embodiment can releasably carry active agents. Modified amino acids can also form drug containing microspheres. These mixtures are suitable for oral administration of biologically active agents to animals. Methods for the preparation of such amino acids are also disclosed.

Description

ORAL DRUG DELIVERY COMPOSITIONS AND METHODS
FIELD OF THE INVENTION

The present invention relates to compositions suitable for oral drug delivery, and in particular to compositions in which modified amino acids and modified amino acid derivatives are used as carriers for sensitive agents such as bioactive peptides and the like. The modified amino acids or derivatives can form non-covalent mixtures with biologically-active agents which are suitable for oral administration to animals. Methods for the preparation and for the administration of such compositions are also disclosed.

BACKGROUND OF THE INVENTION

Conventional means for delivering biologically-active agents, including, but not limited to, pharmaceutical and therapeutic agents, to animals are often severely limited by chemical barriers imposed by the body. Oral delivery of many biologically-active agents would be the route of choice if not for chemical and physico-chemical barriers such as the extreme and varying pH in the gastro-intestinal (GI) tract, exposure to powerful digestive enzymes, and the impermeability of gastro-intestinal membranes to the active agent. Among the numerous agents which are not typically suitable for oral administration are biologically-active peptides such as WO 94/23767 _ ~ ~ 60693 PCT/US94/04560 calcitonin and insulin. Examples of other compounds which are affected by these physico-chemical barriers are polysaccharides and particularly mucopolysaccharides, including, but not limited to, heparin; heparinoids;
antibiotics; and other organic substances. These agents are rapidly destroyed in the gastro-intestinal tract by acid hydrolysis, enzymes, or the like.
Prior methods for orally administering vulnerable pharmacological agents have relied on the co-administration of adjuvants (e.g., resorcinols and non-ionic surfactants such as polyoxyethylene oleyl ether and n-hexadecyl polyethylene ether) to increase artificially the permeability of the intestinal walls; and on the co-administration of enzymatic inhibitors (e.g., pancreatic trypsin inhibitor, diisopropylfluorophosphate (DFF) and trasylol) to inhibit enzymatic degradation. Liposomes have also been described as drug delivery systems for insulin and.
heparin. See, for instance, U.S. Patent No. 4,239,754; Patel et al. (1976) FEBS Letters Vol. 62, page 60; and Hashimoto et al. (1979) Endocrinol. Japan, Vol. 26, page 337.
However, broad spectrum use of the aforementioned drug delivery systems is precluded for reasons including: (1) the need to use toxic amounts of adjuvants or inhibitors; (2) the lack of suitable low MW cargoes; (3) the poor stability and inadequate shelf life of the systems; (4) the difficulties in manufacturing the systems; (5) the failure of the systems to protect the active ingredient; and (6) the failure of the systems to promote absorption of the active agent.
More recently, microspheres of artificial polymers or proteinoids of mixed amino acids have been described for delivery of pharmaceuticals. For example, U.S. Patent No.
4,925,673 describes drug containing microsphere constructs as well as methods for their preparation and use. These proteinoid microspheres are useful for delivery of a number of active agents.
There is still a need in the art for simple and inexpensive delivery systems which are easily prepared and which can deliver a broad range of biologically-active agents.

SUNIlKARY OF THE INVENTION

Compositions for orally delivering biologically active agents incorporating modified amino acids, amino acid derivatives, peptides and peptide derivatives as carriers are provided. These compositions as broadly described hereinafter comprise:
(A) at least one biologically-active agent; and (B) at least one carrier comprising:
(a) (i) at least one acylated aldehyde of an amino acid, (ii) at least one acylated ketone of an amino acid, (iii) at least one acylated aldehyde of a peptide, (iv) at least one acylated ketone of a peptide, or (v) any combination of (a)(i), (a)(ii), (a) (iii) and (a) (iv) ;
(b) (i) carboxymethyl-phenylalamine-leucine, (ii) 2-carboxy-3-phenylpropionyl-leucine, (iii) 2-benzylsuccinic acid, (iv) an actinonin, or (v) a compound having the formula:
Ar-Y- (R1)n-OH

wherein: Ar is a substituted or unsubstituted phenyl or naphthyl;

~
Y is -C- or -S02-;
II
R1 is -N(R4)-R3-C-; wherein:

R3 is C1 to C24 alkyl, C1 to C24 alkenyl, phenyl, naphtyl, (C1 to C1p alkyl) phenyl, (C1 to C10 alkenyl) phenyl, (C1 to C10 alkyl) naphthyl, (C1 to C10 alkenyl) naphtyl, phenyl (C1 to Clp alkyl), phenyl (C1 to C10 alkenyl), naphtyl (C1 to C10 alkyl) and naphtyl (C1 to C10 alkenyl);

R3 is optionally substituted with C1 to C4 alkyl, Cl to C4 alkenyl, C1 to C4 alkoxy, -OH, -SH, -C02R5, cycloalkyl, cycloalkenyl, heterocyclic, aryl, alkaryl, heteroaryl or heteroalkaryl or any combination thereof;

R5 is hydrogen, C1 to C4 alkyl or C1 to C4 alkenyl;

R3 is optionally interrupted by oxygen, nitroven, sulfur or any combination thereof; and R4 is hydrogen, C1 to C4 alkyl or C1 to C4 al.kenyl;
and n is from 1 to about 5;
(vi) or any combination of (i), (b)(ii), (b)(iii), (b)(iv) and (b)(v) or (a) a combination of (a) and (b).

The invention as claimed is however restricted to oral compositions comprising:
(A) at least one biologically-active agent selected from the group consisting of human growth hormone, bovine growth hormone, growth hormone-releasing hormone, an interferon, interleukin-I, interleukin-II, insulin, heparin, low molecular weight heparin, calcitonin, erythropoietin, atrial naturetic factor, an 4a antigen, a monoclonal antibody, somatostatin, adrenocorticotropin, gonadotropin releasing hormone, oxytocin, vasopressin, cromolyn sodium, vancomycin, desferrioxamine and any combination thereof; and (B) a compound having the formula:
Ar-Y- ( Rl ) n-OH
wherein:
Ar is a phenyl or naphthyl optionally substituted with C, to C6 alkyl, Cl to C6 alkenyl, C, to C6 alkoxy, hydroxy, thio, or C02R6 wherein R6 is hydrogen, Cl to C6 alkyl or C, to C6 alkenyl;

Y is -C- or -S02 ;

R1 is -N (R4) -R3-C-; wherein:

R3 is C1 to C24 alkyl, C1 to C24 alkenyl, phenyl, naphthyl, (C1 to. Clo alkyl) phenyl, (C1 to Clo alkenyl) phenyl, (C1 to Clo alkyl) naphthyl, (C1 to Clo alkenyl) naphthyl, phenyl (C1 to Clo alkyl), phenyl (C1 to C10 alkenyl), naphthyl (C1 to Clo alkyl) , or naphthyl (C1 to Clo alkenyl);

R3 is unsubstituted or substituted with C1 to C4 alkyl, C1 to C4 alkenyl, Cl to C4 alkoxy, -OH, -SH, -C02R5 cycloalkyl, cycloalkenyl, heterocyclic, aryl, alkaryl, heteroaryl or heterbalkaryl or any combination thereof;

4b R5 is hydrogen, C1 to C4 alkyl or C1 to C4 alkenyl;
R3 is uninterrupted or interrupted by oxygen, nitrogen, sulfur or any combination thereof; and R4 is hydrogen, C1 to C4 alkyl or C. to C4 alkenyl;
and n is an integer from 1 to 5.

Also contemplated is a method for preparing these compositions which comprises mixing at least one biologically active agent as described above, with at least one carrier as described above, and optionally, a dosage vehicle.

In an alternative embodiment, these non-toxic carriers are orally administered to animals as part of a delivery system by blending or mixing the carriers with a biologically active agent prior to administration. The carriers may also form microspheres in the presence of the active agent. The microspheres' containing the active agent are then orally administered. Also contemplated by the present invention are dosage unit forms that include these compositions. .

DESCRIPTION OF THE DRAWINGS

Figure 1 is a graphic illustration of the results of oral gavage testing in rats using salmon calcitonin with acetyl phenylalanine aldehyde, carbomethoxyPhe-Leu-OH, and acetyl-Phe-Leu-Leu-Arg aldehyde carriers.

Figure 2 is a graphic illustration of the results of oral gavage testing in rats using salmon calcitonin with acetylleucine aldehyde and acetylphenylalanine aldehyde cariers.

WO 94/23767 - 21! o 69" PCT/US94/04560 Figure 3 is a graphic illustration of the results of oral gavage testing in rats using salmon calcitonin with acetylphenylalanine aldehyde and carbomethoxyPhe-Leu-OH
carriers.
5 Figure 4 is a graphic illustration of the results of oral gavage testing in rats using salmon calcitonin with acetylphenylalanine aldehyde, acetylLeu-Leu-Arg aldehyde and carbomethoxyPhe-Leu-OH carriers.
Figure 5 is a graphic illustration of the results of intraduodenal injection testing in rats using salmon calcitonin with acetylphenylalanine aldehyde and 4-(phenylsulfonamido)-4-phenylbutyric acid carriers.
Figure 6 is a graphic illustration of the results of oral gavage testing in rats using salmon calcitonin with acetylphenylalanine aldehyde, N-acetyllysinone, and acetyl-Leu aldehyde carriers.
Figure 7 is a graphic illustration of the results of intraduodenal injection testing in rats using salmon calcitonin with acetylphenylalanine aldehyde carrier in aqueous ethanol, dimethyl sulfoxide (DMSO), and olive oil dosing vehicles, and in a DMSO dosing vehicle alone.
Figure 8 is a graphic illustration of the results of oral gavage testing in rats using salmon calcitonin with cyclohexanoyl-phenylalanine aldehyde carrier.
Figure 9 is a graphic illustration of rat serum calcium levels after oral administration of two dosage levels of a modified amino acid microsphere preparation containing salmon calcitonin and a soluble modified amino acid preparation containing salmon calcitonin after pre-dosing with a sodium bicarbonate solution.
Figure 10 is a graphic illustration of the results of oral gavage testing in rats using salmon calcitonin with = acetyl-Phe aldehyde, actinonin, and carbomethoxy-Phe-Leu-OH
carriers.
Figure 11 is a graphic illustration of the results of oral gavage testing in rats using salmon calcitonin with 4-(phenylsulfonamido)-4-phenylbutyric acid carrier.

SUg~ITUl-E,SHEET (RULE 26) WO 94/23767 - (a 16O 693 PCT/US94/04560 Figure 12 is a graphic illustration of the results of oral gavage testing in rats using salmon calcitonin with 3-(phenylsulfonamido)benzoic acid and 4-(phenylsulfonamido)-hippuric acid carriers.
Figure 13 is a graphic illustration of the results of oral gavage testing in rats using salmon calcitonin with 4-(phenylsulfonamido)-4-phenylbutyric acid and 4-(phenylsulfonamido)benzoic acid carriers.
Figure 14 is a graphic illustration of the results of oral gavage testing in rats using salmon calcitonin with 4-(phenylsulfonamido)-4-phenylbutyric acid and 4-(phenylsulfonamido)phenylacetic acid carriers.
Figure 15 is a graphic illustration of the results of oral gavage testing in rats using interferon a2b (rhiFN) with 4-(phenylsulfonamido)-4-phenylbutyric acid carrier.
Figure 16 is a graphic illustration of the results of oral gavage testing in rats using interferon cx2b with 4-(phenylsulfonamidomethyl)benzoic acid carrier.
Figure 17 is a graphic illustration of the results of oral gavage testing in rats using interferon cx2b with 4-(phenylsulfonamido)phenylacetic acid as carrier.
Figure 18 is a graphic illustration of the results of oral gavage testing in rats using interferon aa2b with 4-(phenylsulfonamido)hippuric acid carrier..
Figures 19 and 20 are graphic illustrations of the results of oral gavage testing in hypophysectomized rats using growth hormone alone and at two dosage levels with 4-(phenylsulfonamido)-4-phenylbutyric acid carrier.
Figure 21 is a graphic illustration of the results of oral gavage testing in normal rats using growth hormone with 4-(phenylsulfonamido)-4-phenylbutyric acid carrier.
Figure 22 is a graphic illustration of the results of oral gavage testing in rats using disodium cromoglycate =
with 4-(phenylsulfonamido)-4-phenylbutyric acid as carrier.
Detailed Description of the Invention Amino acids and amino acid derivatives, in modified form, may be used to deliver orally sensitive biologically-active agents, including, but not limited to, hormones such as calcitonin, insulin, and polysaccharides such as heparin, which would not be considered orally administrable for various reasons. Insulin, for example is sensitive to the denaturing conditions of the gastro--intestinal (GI) tract. Also, heparin, by virtue of its charge and hydrophilic nature, is not readily absorbed from the gastro-intestinal tract. In contrast to the modified amino acids and modified amino acid derivatives of the present invention, unmodified free amino acids do not provide protection against degradation in the GI tract for labile bioactive agents.
The compositions of the subject invention are useful for administering biologically-active agents to any animals such as birds; mammals, such as primates and particularly humans; and insects.
Other advantages provided by the present invention include the use of readily available and inexpensive starting materials in cost-effective methods for preparing and isolating modified amino acid derivatives. These methods are simple to perform and are amenable to industrial scale-up for commercial production.
Biologically-active agents suitable for use with carriers disclosed herein include, but are not limited to, peptides, and particularly small peptide hormones, which by themselves do not pass or only pass slowly through the gastro-intestinal mucosa and/or are susceptible to chemical cleavage by acids and enzymes in the gastro-intestinal tract; polysaccharides and particularly mixtures of muco-polysaccharides; carbohydrates; lipids; or any combination thereof. Examples include, but are not limited to, human growth hormone; bovine growth hormone; growth hormone releasing hormone; interferons; interleukin-I; insulin;
heparin, and particularly low molecular weight heparin;
= 35 calcitonin; erythropoietin; atrial naturetic factor;
antigens; monoclonal antibodies; somatostatin;
adrenocorticotropin; gonadotropin releasing hormone;
oxytocin; vasopressin; cromolyn sodium (sodium or disodium SUBSTITUTE SHEET (RULE 26) WO 94/23767 ej a 6 9 3 PCT/US94/04560 cromoglycate); vancomycin; desferrioxamine (DFO); or any combination thereof.
Additionally the carriers of the present invention can be used to deliver other active agents such as pesticides and the like.
The term amino acid as used herein includes any carboxylic acid having at least one free amine group including naturally occurring and synthetic amino acids.
The preferred amino acids are a-amino acids, and preferably are naturally occurring a-amino acids although non-cx-amino acids are useful as well.
Poly amino acids as used herein refers to peptides or two or more amino acids linked by a bond formed by other groups which can be linked, e.g., an ester, anhydride or an anhydride linkage.
The term peptide is meant to include two or more amino acids joined by a peptide bond. Peptides can vary in length from dipeptides with 2 to poly peptides with several hundred amino acids. See Chambers Biological Dictionary, editor Peter M. B. Walker, Cambridge, England: Chambers Cambridge, 1989, page 215. The peptides most useful in the practice of the present invention include di-peptides, tri-peptides, tetra-peptides, and penta-peptides. The preferred peptides are di-peptides, tri-peptides. Peptides can be homo- or hetero- peptides and can include natural amino acids, synthetic amino acids, or any combination thereof.
The term amino acid derivatives and peptide derivatives as used herein are meant to include amino acid aldehydes or ketones and/or peptide aldehydes or ketones where the -COOH group has been converted to a ketone or aldehyde.
The terms modified amino acids, peptides, and derivatives thereof are meant to include amino acids, amino acid derivatives, peptides and peptide derivatives which have been modified as described below by acylating or sulfonating at least one free amine group, with an acylating SUBSTITUTE SHEET (RULE 26) WO 94/23767 216a 693 PCT/US94/04560 or sulfonating agent which reacts with at least one of the free amine groups present.
The preferred naturally occurring amino acids for . use in the present invention as amino acids or components of a peptide are alanine, arginine, asparagine, aspartic acid, , citrulline, cysteine, cystine, glutamine, glycine, histidine, isoleucine, leucine, lysine, methionine, ornithine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine, valine, hydroxy proline, y-carboxyglutamate, or 0-phosphoserine. The most preferred amino acids are arginine, leucine, lysine, phenylalanine, tyrosine and valine.
The preferred non-naturally occurring amino acids for use in the present invention as amino acids or components of a peptide are fl-alanine, phenyiglycine, a-aminobutyric acid, ry-amino butyric acid, 4-(4-aminophenyl)butyric acid, cx-amino isobutyric acid, e-aminocaproic acid, 7-aminoheptanoic acid, (3-aspartic acid, aminobenzoic acid, (aminomethyl)benzoic acid, aminophenylacetic acid, aminohippuric acid, 7-glutamic acid, cysteine(ACM), E-lysine, E-lysine (A-Fmoc), methionine sulfone, norleucine, norvaline, ornithine, d-ornithine, p-nitrophenylalanine, hydroxy proline, and thioproline.
The amino acids useful in the practice of the subject invention have the formula:
HN (R4) - (R2) n-OH

RZ has the formula -R3 -C- wherein R3 is CI to C~
alkyl, C1 to C24 alkenyl, phenyl, naphthyl, (Cl to Clo alkyl )-phenyl phC1 to Cto alkenyl ) phenyl , ( C1 to Clo alkyl ) naphthyl , (C, to CIO alkenyl ) naphthyl, phenyl ( CI to Clo alkyl ), phenyl ( C1 to Clo alkenyl ), naphthyl ( CI to Clo alkyl) and naphthyl ( C1 to Cio alkenyl) ;
optionally R3 is substituted with Cl to C4 alkyl, C1 to C4 alkenyl, Cl to C4 alkoxy, -OH, -SH and - COZRs or any combination thereof;
RS is hydrogen, C, to C4 alkyl or Cl to C4 alkenyl ;

R3 is optionally interrupted by oxygen, nitrogen, sulfur or any combination thereof; and R4 is hydrogen, C, to C4 alkyl or C1 to C4 alkenyl.
The phenyl or naphthyl groups can be optionally 5 substituted. Suitable but non-limiting examples of substitutents are C1 to C6 alkyl, Cl to C6 alkenyl, alkoxy having from 1 to 6 carbon atoms, hydroxy, thio, or C02R6 wherein R6 is hydrogen, Cl to C6 alkyl, C1 to C6 alkenyl.
The amino acid derivatives or peptide derivatives 10 of the present invention can be readily prepared by reduction of amino acid esters or peptide esters with an appropriate reducing agent. For example, amino acid aldehydes or peptide aldehydes can be prepared as described in an article by R. Chen et al., Biochemistry, 1979, 18, 921-926. Amino acid or peptide ketones can be prepared by the procedure described in Organic Syntheses, Col. Vol. IV, Wiley, (1963), pages 5-6. Amino acids, peptides, amino acid"
esters, peptide esters, and other necessary reagents to prepare these derivatives are readily available from a number of commercial sources such as Aldrich Chemical Co.
(Milwaukee, WI, USA); Sigma. Chemical Co. (St. Louis, MO, USA); and Fluka Chemical Corp. (Ronkonkoma, NY, USA).
The amino acids and peptides are modified by acylating or sulfonating at least one free amine group, with an acylating or sulfonating agent which reacts with at least one of the free amine groups present. Suitable, but non-limiting, examples of agents useful for modifying amino acids or peptides useful in practicing the present invention include acylating and sulfonating agents having the formula R' ( X
or R7--SOr-X wherein R7 is alkyl or alkenyl, preferably having from 1 to 20 carbon atoms, or aromatic preferably having from 6 to 20 carbon atoms.
The R7 group can be substituted or unsubstituted, The n :ferred substitutents include Cl to C4 alkyl, C1 to C4 alkenyl, C1 to C4 alkoxy, COZRs wherein Rg is hydrogen, Cl to C4 alkyl or Cl to C4 alkenyl.
Preferably, R' is methyl, ethyl, phenyl, benzyl or naphthyl. More preferably, R7 is phenyl, or acetyl. X is a leaving group. In a reaction in which the substrate = molecule becomes cleaved, part of it (the part not containing the carbon) is usually called the leaving group.
. See Advanced Organic Chemistry, 2d edition, Jerry March, New York: McGraw-Hill Book (1977), page 187, Typical leaving groups include, but are not limited to, halogens such as chlorine, bromine and iodine.
Examples of the acylating and sulfonating agents for amino acids and peptides include, but are not limited to, acyl halides such as acetyl chloride, propyl chloride, benzoyl chloride, hippuryl chloride and the like; sulfonyl halides such as b-enzene sulfonyl chloride, and anhydrides, such as acetic anhydride, propyl anhydride, benzoic anhydride, hippuric anhydride and the like. The preferred acylating and sulfonating agents are benzoyl chloride, benzene sulfonyl chloride, and hippuryl chloride.
The modified acid compounds have the formula:
Ar-Y- ( R1) -OH
wherein Ar is a substituted or unsubstituted phenyl or naphthyl;
O
Y is A_ or -SOr-, R' has the formula -V (R4)-3.23-R-, wherein:
R3 is Cl to Cu alkyl, Ci to C24 alkenyl, phenyl, naphthyl,( C1 to Clo alkyl) phenyl,( Cl to Clo alkenyl) phenyl, ( C1 to Clo alkyl) naphthyl,( C1 to Clo alkenyl) naphthyl, phenyl (Cl to Cio alkyl ), phenyl (Cl to CIO alkenyl ), naphthyl ( C1 to Clo alkyl) and naphthyl ( Ci to Cio alkenyl );
R3 is optionally substituted with Cl to C4 alkyl, C, to C4 alkenyl, Cl to C4 alkoxy, -OH, -SH and -C02R5 or any combination thereof;
= R5 is hydrogen, C, to C4 alkyl or Cl to C4 alkenyl;
R3 is optionally interrupted by oxygen, nitrogen, sulfur or any combination thereof; and R4 is hydrogen, C, to C4 alkyl or Cl to C4 alkenyl.
The amino acid derivatives and peptide derivatives are modified by acylating at least one free amine group, WO 94/23767 _ 2160693 PCT/US94/04560 with an acylating agent which reacts with at least one of the free amine groups present. Suitable, but non-limiting, examples of acylating agents useful for modifying amino acid derivatives and peptide derivatives useful in practicing the present invention include acid chloride acylating agents having the formula R9-CX or wherein:
R9 is alkyl or alkenyl, preferably having from 1 to 20 carbon atoms, cycloalkyl or cycloalkenyl, preferably having from 1 to 20 carbon atoms, or aromatic preferably having from 6 to 20 carbon atoms. The R9 group can be substituted or unsubstituted, The preferred substituents include Cl to C4 alkyl, Ci to C4 alkenyl, Cl to C4 alkoxy, CO2R10 wherein R10 is hydrogen, Cl to C4 alkyl or Cl to C4 alkenyl.
Preferably, R9 is methyl, ethyl, cyclohexyl, cyclopentyl, cycloheptyl, phenyl, benzyl or naphthyl. More preferably, R9 is phenyl, cyclohexyl cyclopentyl, cycloheptyl, or acetyl.
X is a leaving group. Typical leaving groups include, but are not limited to, halogens such as chlorine, bromine and iodine.
Examples of the acylating agents for amino acid derivatives and peptide derivatives include, but are not limited to, acyl halides such as acetyl chloride, propyl chloride, cyclohexanoyl chloride, cyclopentanoyl chloride, and cycloheptanoyl chloride, benzoyl chloride, hippuryl chloride and the like; and anhydrides, such as acetic anhydride, propyl anhydride, cyclohexanoic anhydride, benzoic anhydride, hippuric anhydride and the like. The preferred acylating agents are benzoyl chloride, benzene sulfonyl chloride, hippuryl chloride, acetyl chloride, cyclohexanoyl chloride, cyclopentanoyl chloride, and cycloheptanoyl chloride.
The amine groups can also be modified by the reaction of a carboxylic acid with coupling agents such as dicyclohexylcarbodiimide and the like. In a peptide one or more of the amino acids may be derivatized (an aldehyde or a ketone) and/or modified (acylated).
SUBSTITUTE SHEET (RULE 26) WO 94/23767 216" 693 PCT/US94/04560 Also suitable as a carrier alone or in combination with the modified amino acid or peptide derivatives are the carbomethoxy modified amino acids carboxy-methyl-phenylalanine-leucine, 2-carboxy-3-phenylpropionyl-leucine, 2-benzylsuccinic acid and an actinonin. The actinonin compounds include actinonin or epiactinonin and derivatives thereof. These compounds have the formulas below:

Me Me ~ ~ O Me~Me O
N
N NHOH N` NHOH
~ O H
H O H HOJ O

Me Me Actinonin Epiactinonin Derivatives of these compounds are disclosed in U.S. Patent No. 5,206,384.
Actinonin derivatives have the formula:

Rt? C
, CHZ H 0 N N

CH3/ CH3 R1i wherein R11 is sulfoxymethyl or carboxyl or a substituted carboxy group selected from carboxamide, hydroxyaminocarbonyl and alkoxycarbonyl groups; and R12 is hydroxyl, alkoxy, hydroxyamino or sulfoxyamino group.
The modified amino acid derivatives or peptide derivatives can be readily prepared and modified by methods known to t-iia,se skilled in the art. For example, the modified amino acid derivatives of the present invention may be prepared by reacting a single amino acid derivative or peptide derivative or mixtures of two or more amino acid or peptide derivatives, with an acylating agent or an amine modifying agent which reacts with free amino moieties present in the derivatives to form amides. The amino acid or peptide can be modified and subsequently derivatized, derivatized and subsequently ^odified, or simultaneously WO 94/23767 ^ ~ ~ ~ 0693 PCT/US94/04560 modified and derivatized. Protecting groups may be used to avoid unwanted side reactions as would be known to those skilled in the art.
The modified amino acids and modified amino acid derivatives of the present invention may also be prepared by reacting single amino acids, mixtures of two or more kinds of amino acids, or amino acid esters with an amine modifying agent which reacts with free amino moieties present in the amino acids to form amides or sulfonamides. Amino acids and amino acid esters are readily available from a number of commercial sources such as Aldrich Chemical Co. (Milwaukee, WI, USA); Sigma Chemical Co. (St. Louis, MO, USA); and Fluka Chemical Corp. (Ronkonkoma, NY, USA).
For example, the amino acids can be dissolved in aqueous alkaline solution of a metal hydroxide, e.g., sodium or potassium hydroxide, and heated at a temperature ranging between about 5 C and about 70 C, preferably between about 10 C and about 40 C, for a period ranging between about 1 hour and about 4 hours, preferably about 2.5 hours. The amount of alkali employed per equivalent of NHZ groups in the amino acids generally ranges between about 1.25 and about 3 mmole, preferably between about 1.5.and about 2.25 mmole per equivalent of NH2. The pH of the solution generally ranges between about 8 and about 13, preferably ranging between about 10 and about 12.
Thereafter, an amino modifying agent is added to the amino acid solution while stirring. The temperature of the mixture is maintained at a temperature generally ranging between about 5 C and about 70 C, preferably between about 10 C and about 40 C, for a period ranging between about 1 and about 4 hours. The amount of amino modifying agent employed in relation to the quantity of amino acids is based on the moles of total free NH, in the amino acids. In general, the amino modifying agent is employed in an amount ranging between about 0.5 and about 2.5 mole equivalents, preferably between about 0.75 and about 1.25 equivalents, per molar equivalent of total NH2 groups in the amino acids.

The reaction is quenched by adjusting the pH of the mixture with a suitable acid, e.g., concentrated hydrochloric acid, until the pH reaches between about 2 and about 3. The mixture separates on standing at room 5 temperature to form a transparent upper layer and a white or off-white precipitate. The upper layer is discarded and modified amino acids are collected from the lower layer by filtration or decantation. The crude modified amino acids are then dissolved in water at a pH ranging between about 9 10 and about 13, preferably between about 11 and about 13.
Insoluble materials are removed by filtration and the filtrate is dried in vacuo. The yield of modified amino acids generally ranges between about 30 and about 60%-, and usually about 45%-.
15 If desired, amino acid esters, such as, for example methyl or ethyl esters of amino acids, may be used to prepare the modified amino acids of the invention. The amino acid esters, dissolved in a suitable organic solvent such as dimethylformamide or pyridine, are reacted with the amino modifying agent at a temperature ranging between about 5 C and about 70 C, preferably about 25 C, for a period ranging between about 7 and about 24 hours. The amount of amino modifying agents used relative to the amino acid esters are the same as described above for amino acids.
Thereafter, the reaction solvent is removed under negative pressure and the ester functionality is removed by hydrolyzing the modified amino acid ester with a suitable alkaline solution, e.g. iN sodium hydroxide, at a temperature ranging between about 50 C and about 80 C, preferably about 70 C, for a period of time sufficient to hydrolyze off the ester group and form the modified amino acid having a free carboxyl group. The hydrolysis mixture . is then cooled to room temperature and acidified, e.g.
aqueous 25W hydrochloric acid solution, to a pH ranging between about 2 and about 2.5. The modified amino acid precipitates out of solution and is recovered by conventional means such as filtration or decantation.

SUBSTITUTE SHEET (RULE 26) WO 94/23767 21~i L* 0U `a 9ca PCT/US94/04560 16 y'S

The modified amino acids may be purified by recrystallization or by fractionation on solid column supports. Suitable recrystallization solvent systems include acetonitrile, methanol and tetrahydrofuran.
Fractionation may be performed on a suitable solid column supports such as alumina, using methanol/n-propanol mixtures as the mobile phase; reverse phase column supports using trifluoroacetic acid/acetonitrile mixtures as the mobile phase; and ion exchange chromatography using water as the mobile phase. When anion exchange chromatography is performed, a subsequent 0-500 mM sodium chloride gradient is employed. The modified amino acids may also be purified by extraction with a lower alcohol such as methanol, butanol, or isopropanol to remove low molecular weight non-sphere making material.
Suitable modified amino acid derivatives include, but are not limited to, N-cyclohexanoyl-Phe aldehyde, N-acetyl-Phe-aldehyde, N-acetyl-Tyr ketone, N-acetyl-Lys ketone and N-acetyl-Leu ketone. Special mention is made of the modified amino acid derivative N-cyclohexanoyl phenylalanine aldehyde.
Special mention is made of compositions in which the biologically-active agent includes, calcitonin and the carrier includes acetyl phenylalanine aldehyde, carbomethoxy phenylalanylleucine and acetyl-Phe-Leu-Leu aldehyde.
Special mention is also made of a composition which includes 1.5 Ag/ml of the biologically-active agent calcitonin and the carrier includes 132 mg/ml of acetyl phenylalanine, 33 mg/ml of carbomethoxy phenylalanylleucine, and 25 mg/ml of acetyl-Phe-Leu-Leu-Arg aldehyde.
In one embodiment, the modified and/or modified derivatized amino acids may be used directly as a delivery carrier by simply mixing the carrier with the active ingredient prior to administration. In an alternative embodiment, the modified amino acids may be used to form microspheres containing the active agent. The modified and/or modified derivatized amino acids of the invention are-particularly useful for the oral administration of certain pharmacological agents, e.g., small peptide hormones, which, by themselves, do not pass or only pass slowly through the gastro-intestinal mucosa and/or are susceptible to chemical cleavage by acids and enzymes in the gastro-intestinal tract.
If the modified amino acids are to be converted into microspheres, the mixture is optionally heated to a temperature ranging between about 20 and about 50 C, preferably about 40 C, until the modified amino acid(s) dissolve. The final solution contains between from about 1 mg and about 2000 mg of modified amino acids per mL of solution, preferably between about 1 and about 500 mg per mL. The concentration of active agent in the final solution varies and is dependent on the required dosage for treatment. When necessary, the exact concentration can be determined by, for example, reverse phase HPLC analysis.
When the modified amino acids are used to prepare microspheres, another useful procedure is as follows:
Modified amino acids are dissolved in deionized water at a concentration ranging between about 75 and about 200 mg/ml, preferably about 100 mg/mi at a temperature between about C and about 60 C, preferably about 40 C. Particulate matter remaining in the solution may be removed by conventional means such as filtration.
25 Thereafter, the modified amino acid solution, maintained at a temperature of about 40 C, is mixed 1:1 (V/V) with an aqueous acid solution (also at about 40 C) having an acid concentration ranging between about 0.05 N
and about 2 N, preferably about 1.7 N. The resulting mixture is further incubated at 40 C for a period of time effective for microsphere formation, as observed by light microscopy. In practicing this invention, the preferred order of addition is to add the modified amino acid solution to the aqueous acid solution.
Suitable acids for microsphere formation include any acid which does not 18 + 216~fn 693 PCT/US94/04560 (a) adversely effect the modified amino acids, e.g., initiate or propagate chemical decomposition;
(b) interfere with microsphere formation;
(c) interfere with microsphere encapsulation of the cargo; and (d) adversely interact with the cargo.
Preferred acids for use in this invention include acetic acid, citric acid, hydrochloric acid, phosphoric acid, malic acid and maleic acid.
In practicing the invention, a microsphere stabilizing additive may be incorporated into the'aqueous acid solution or into the amino acid solution prior to the microsphere formation process. With some drugs the presence of such additives promotes the stability and/or dispersibility of the microspheres in solution.
The stabilizing additives may be employed at a concentration ranging between about 0.1 and 5k (w/v), preferably about 0.5 k (w/v). Suitable, but non-limiting, examples of microsphere stabilizing additives include gum acacia, gelatin, methyl cellulose, polyethylene glycol, and polylysine. The preferred stabilizing additives are gum acacia, gelatin and methyl cellulose.
Under the above conditions, the modified amino acid molecules form hollow or solid matrix type microspheres wherein the cargo is distributed in a carrier matrix or capsule type microspheres encapsulating liquid or solid cargo. If the modified amino acid microspheres are formed in the presence of a soluble material, e.g., a pharmaceutital agent in the aforementioned aqueous acid solution, this material will be encapsulated within the microspheres. In this way, one can encapsulate pharmacologically active materials such as peptides, proteins, and polysaccharides as well as charged organic molecules, e.g., antimicrobial agents, which normally have poor bioavailability by the oral route. The amount of pharmaceutical agent which may be encapsulated by the microsphere is dependent on a number of factors which lg 2160693 include the concentration of agent in the encapsulating solution, as well as the affinity of the cargo for the carrier.
The modified amino acid microspheres of the invention are pharmacologically harmless and do not alter the physiological and biological properties of the active agent. Furthermore, the encapsulation process does not alter the pharmacological properties of the active agent.
Any pharmacological agent can be encapsulated within the amino acid microspheres. The system is particularly advantageous for delivering chemical or biological agents which otherwise would be destroyed or rendered less effective by conditions encountered within the body of the animal to which it is administered, before the microsphere reaches its target zone (i.e., the area in which the contents of the microsphere are to be released) and pharmacological agents which are poorly absorbed in the gastro-intestinal tract. The target zones can vary depending upon the drug employed.
The particle size of the microsphere plays an important role in determining release of the active agent in the targeted area of.the gastro-intestinal tract. The preferred microspheres have diameters between about < 0.1 microns and about 10 microns, preferably between about 0.5 microns and about 5 microns. The microspheres are sufficiently small to release effectively the active agent at the targeted area within the gastro-intestinal tract.
Small microspheres can also be administered parenterally by being suspended in an appropriate carrier fluid (e.g., isotonic saline) and injected directly into the circulatory system, intramuscularly or subcutaneously. The mode of administration selected will vary, of course, depending upon the requirement of the active agent being administered.
Large amino acid microspheres (>50 microns) tend to be less effective as oral delivery systems.
The size of the microspheres formed by contacting modified amino acid with water or an aqueous solution containing active agents can be controlled by manipulating a WO 94/23767 20 -iv c) qAta4!(jn n n~j`~,f3 PCTIUS94/04560 variety of physical or chemical parameters, such as the pH, osmolarity or ionic strength of the encapsulating solution, size of the ions in solution and by the choice of acid used in the encapsulating process.
Typically, the pharmacological compositions of the present invention are prepared by mixing an aqueous solution of the carrier with an aqueous solution of the active ingredient, just prior to administration. Alternatively, the carrier and biologically active ingredient can be admixed during the manufacturing process. The solutions may optionally contain additives such as phosphate buffer salts, citric acid, acetic acid, gelatin and gum acacia.
In practicing the invention, stabilizing additives may be incorporated into the carrier solution. With some drugs, the presence of such additives promotes the stability and dispersibility of the agent in solution.
The stabilizing additives may be employed at a concentration ranging between about 0.1 and 5!~ (W/V), preferably about 0.5 %- (W/V). Suitable, but non-limiting, examples of stabilizing additives include gum acacia, gelatin, methyl cellulose, polyethylene glycol, and polylysine. The preferred stabilizing additives are gum acacia, gelatin and methyl cellulose.
The amount of active agent in the composition typically is a pharmacologically or biologically effective amount. However, the amount can be less than a pharmacologically or biologically effective amount when the composition is used in a dosage unit form, such as a capsule, a tablet or a liquid, because the dosage unit form may contain a multiplicity of carrier/biologically-active agent compositions or may contain a divided pharmacologically or biologically effective amount. The total effective amounts will be administered by cumulative units containing in total pharmacologically or biologically active amounts of biologically-active agent.
The total amount of biologically-active agent to be used can be determined by those skilled in the art.
However, it has surprisingly been found that with certain SUBSTITUTE SHEET (RULE 26) biologically-active agents, such as calcitonin, the use of the presently disclosed carriers provides extremely efficient delivery. Therefore, lower amounts of biologically-active agent than those used in prior dosage unit forms or delivery systems can be administered to the subject, while still achieving the same blood levels and therapeutic effects.
The amount of carrier in the present composition is a delivery effective amount and can be determined for any particular carrier or biologically-active agent by methods known to those skilled in the art.
Dosage unit forms can also include any of excipients; diluents; disintegrants; lubricants;
plasticizers; colorants; and dosing vehicles, including, but not limited to water, 1,2-propane diol, ethanol, olive oil, or any combination thereof.
Administration of the present compositions or dosage unit forms is oral or by intraduodenal injection.
EXAMPLES
The invention will now be illustrated in the following non-limiting examples which are illustrative of the invention but are not intended to limit the scope of the invention.

PREPARATION OF N-CYCLOHEXANOYLPHENYLALANINE ALDEHYDE:
Phenylalanine methyl ester (1 g., 0.0046 moles) was dissolved in pyridine 5 mL. Cyclohexanoyl chloride (0.62 mL) was added and the mixture was stirred for 2 hours.
The reaction mixture was poured onto hydrochloric acid (1N) and crushed ice. The aqueous mixture was extracted twice with toluene. The combined toluene extracts were concentrated in vacuo to give 1.1 g of crude N-cyclohexan-oylphenylalanine methyl ester.
N-Cyclohexanoylphenylalanine methyl ester (0.5 g) was dissolved in ethylene glycol dimethyl ether (20 mL).
The solution was cooled to -70 C and diisobutylaluminum hydride (2.04 mL of a 1.5M solution in toluene) was added.
The resulting reaction mixture was stirred at -70 C for 2 hours. The reaction was quenched by dropwise addition of 2N
hydrochloric acid. The mixture was extracted with cold ethyl acetate. The ethyl acetate solution was washed with brine and dried over sodium sulfate. Concentration in vacuo furnished a white solid which was purified by silica gel chromatography. 1H NMR(300 MHz, DMSO-d6): 9.5 (s, 1H), 8.2 (d, 1H) , 7.2 (m, 5H), 4.2 (m, 1H), 3.2 (d, 1H), 2.7 (d, 1H) , 2.1 (m, 1H), 1.6 (br. m, 4H), 1.2 (br. m, 6H).
IR (KBr): 3300, 3050, 2900, 2850, 2800, 1700, 1600, 1500 cm-Mass Spec.: M+1 m/e 261.

PREPARATION OF N-ACETYLPHENYLALANINE ALDEHYDE:
N-Acetylphenylalanine methyl ester (4.2 g, 19 mmol) was dissolved in ethylene glycol dimethyl ether. The solution was cooled to -70 C and diisobutylaluminum hydride (25.3 mL of a 1.5M solution in toluene, 39 mmol) was added.
The resulting reaction mixture was stirred at -70 C for 2 hours. The reaction was quenched by addition of 2N
hydrochloric acid. The mixture was extracted 4 times with cold ethyl acetate and 4 times with toluene. The extracts were combined, washed with brine and dried over magnesium sulfate. Concentration in vacuo followed by silica gel chromatography furnished 2.7 g of a white solid. The NMR
was identical to that reported in the literature, Biochemistry, 1979, 18, 921-926.

PREPARATION OF 3-ACETAMIDO-4-(p-HYDROXY)PHENYL-2-BUTANONE
(N-ACETYLTYROSINONE):
A mixture of tyrosine (28.9, g, 16 mmoi), acetic anhydride (97.9 g,96 mmol) and pyridine (35g, 16 mmol) were heated to 100 C for 1 hour. The reaction mixture was concentrated in vacuo to furnish a yellow oil. The oil was distilled at reduced pressure to furnish 29.9 g or an oil.
SUBSIITUTE SHEET (RULE 26) WO 94/23767 23 - 2160693 P'CT/US94/04560 'H NMR (DMSO-d6): NMR (d6-DMSO); 8.2 (d, 1H), 7.3 (d, 2H), 7.0 (d, 2H), 4.4 (m, 1H), 3.1 (dd, 1H), 2.7 (dd, 1H), 2.3 (s, 3H) , 1.8 (s, 3H) (N-ACETYLLYSINONE):
Following the procedure of Example 3 lysine was converted to N-acetyllysinone.
'H NMR (DMSO-d6): 8.1 (d, 1H), 7.8 (br.m. 1H), 4.1 (m, 1H), 3.0 (m, 2H), 2.0 (s, 3H), 1.9 (s,3H) and 1.3 (br.m, 6H).

(N-ACETYLLEUCINONE):
Following the procedure of Example 3 leucine was converted to N-acetylleucinone.
'H NMR (DMSO-d6): 8.1 (d, 1H), 4.2 (m, 1H), 2.0 (s, 3H), 1.8 (s, 3H) , 0. 8 (d, 6H) .

MODIFICATION OF 4-(4-AMINOPHENYL)BUTYRIC ACID USING BENZENE
SULFONYL CHLORIDE
4-(4-Aminophenyl)butyric acid, (20 g 0.11 moles) was dissolved in 110 mL of aqueous 2N sodium hydroxide solution. After stirring for about 5 minutes at room temperature, benzene sulfonyl chloride (14.2 mL, 0.11 moles) was added dropwise into the amino acid solution over a 15 minute period. After stirring for about 3 hours at room temperatuLerthe mixture was acidified to pH 2 by addition of hydrochloric acid. This furnished a light brown precipitate which was isolated by filtration. The precipitate was washed with warm water and dried. The yield of 4-(phenyl-sulfonamido)4-phenylbutyric acid was 24.3 g (69%). The melting point was 123-25 C.
If necessary, the modified amino acids can be purified by recrystallization and/or chromatography.

CHLORIDE
Following the procedure of Example 6 4-aminobenzoic acid was converted to 4-(phenylsulfonamido)benzoic acid.

MODIFICATION OF 4-AMINOPHENYLACETIC ACID, 4-AMINOHIPPURIC
ACID, AND 4-AMINOMETHYLBENZOIC ACID USING BENZENE SULFONYL
CHLORIDE
Following the procedure of Example 6, 4-aminophenylacetic acid, 4-aminohippuric acid, and 4-amino-methylbenzoic acid were converted to 4-(phenylsulfonamido)-phenylacetic acid, 4-(phenylsulfonamido)hippuric acid, and 4-(phenylsulfonamidomethyl)benzoic acid respectively.

MODIFICATION OF AMINO ACIDS WITH BENZENE SULFONYL CHLORIDE
A mixture of sixteen amino acids were prepared prior to chemical modification. The constituents of the mixture are summarized in Table 1. 65 grams of the amino acid mixture (total concentration of [-NH2] groups = 0.61 moles) was dissolved in 760 mL of 1N sodium hydroxide solution (0.7625 equivalents) at room temperature. After stirring for 20 minutes, benzene sulfonyl chloride (78 ml, 1 eq.) was added over a 20 minute period. The reaction mixture was then stirred for 2.5 hours, without heating. As some precipitation had occurred, additional NaOH solution (2N) was added to the solution until it reached pH 9.3.
The reaction mixture stirred overnight at room temperature.
Thereafter, the mixture was acidified using dilute hydrochloric acid (38!k, 1:4) and a cream colored material precipitated out. The resulting precipitate was isolated by decantation and dissolved in sodium hydroxide (2N). This solution was then reduced in vacuo to give a yellow solid, which was dried on the lyophilizer.

~ WO 94/23767 2.160.~'~ 93 PCT/US94/04560 TABLE 1: Amino Acid Composition No. of moles 5 Weight t of Total of each Amino No. of Moles Amino Acid (g) Weight Acid (x10-2) of -[ - NHZ]
Thr 2.47 3.8 2.07 2.07 Ser 2.25 3.46 2.1 2.1 Ala 4.61 7.1 5.17 5.17 10 Val 4.39 6.76 3.75 3.75 Met 0.53 0.82 0.35 0.35 Ile 2.47 3.8 0.36 0.36 Leu 3.86 5.94 2.95 2.95 Tyr 1.03 1.58 0.56 0.56 15 Phe 4.39 6.76 0.27 0.27 His 2.47 3.8 1.6 3.2 Lys 4.94 7.6 3.4 6.8 Arg 5.13 7.9 2.95 5.90 Glutamine 9.87 15.18 6.76 13.42 20 Glutamic Acid 9.87 15.18 6.70 6.70 Asparagine 3.32 5.11 2.51 5.02 Aspartic Acid 3.32 5.11 2.50 2.50 MODIFICATION OF A MIXTURE OF FIVE AMINO ACIDS USING BENZENE
SULFONYL CHLORIDE
An 86.1g (0.85 moles of NH2) mixture of amino acids (see Table 2) was dissolved in 643 mL (1.5 eq.) of aqueous 2N sodium hydroxide solution. After stirring for 30 minutes at room temperature, benzene sulfonyl chloride (108 mL, 0.86 moles) was added portionwise into the amino acid solution over a 15 minute period. After stirring for 2.5 hours at room temperature, the pH of the reaction mixture (pH 5) was adjusted to pH 9 with additional 2N sodium hydroxide solution. The reaction mixture stirred overnight at room temperature. Thereafter, the pH of the reaction mixture was SUBSTITUTE SHEET (RULE 26) =
WO 94/23767 r 216Q U 9 3 PCTIUS94/04560 adjusted to pH 2.5 by addition of dilute aqueous hydrochloric acid solution (4:1, H20:HC1) and a precipitate of modified amino acids formed. The upper layer was discarded and the resulting yellow precipitate was isolated by decantation, washed with water and dissolved in 2N sodium hydroxide (2N). The solution was reduced in vacuo to give a yellow solid which was lyophilized overnight. The yield of crude modified amino acid was 137.9 g.

Table 2 Amino Acid Moles of Amino Moles of Acid [ -NFI2] x10-2 (x 10"2) Valine 7.5 7.5 Leucine 10.7 10.5 Phenylalanine 13.4 13.4 Lysine 21.0 42.0 Arginine 6.0 12.0 SU$STITUTE SHEET (RULE 26) ~ ~ ~/1~~p3 PCT/US94/04560 WO 94/23767 27 .. !1 +~

MODIFICATION OF A MIXTURE OF FIVE AMINO ACIDS USING BENZOYL
CHLORIDE
An 86 g (0.85 moles of NH2) mixture of amino acids (see Table 2 in Example 10) was dissolved in 637 mL (1.5 eq.) of aqueous 2N sodium hydroxide solution. After stirring for 10 minutes at room temperature, benzoyl chloride (99 mL, 0.85 moles) was added portionwise into the amino acid solution over a 10 minute period. After stirring for 2.5 hours at room temperature, the pH of the reaction mixture (pH 12) was adjusted to pH 2.5 using dilute hydrochloric acid (4:1, H2O:HC1) and a precipitate of modified amino acids formed. After settling for 1 hour, the resulting precipitate was isolated by decantation, washed with water and dissolved in sodium hydroxide (2N). This solution was then reduced in vacuo to give crude modified amino acids as a white solid (220.5.g).

MODIFICATION OF L-VALINE USING BENZENE SULFONYL CHLORIDE
L-Valine (50 g, 0.43 mol) was dissolved in 376 mL
(0.75 eq.) of aqueous 2N sodium hydroxide by stirring at room temperature for 10 minutes. Benzene sulfonyl chloride (68.7 mL, 0.38 mol, 1.25 eq.) was then added to the amino acid solution over a 20 minute period at room temperature.
After stirring for 2 hours at room temperature, a precipitate appeared. The precipitate was dissolved by adding 200 mL of additional 2N sodium hydroxide solution.
After stirring for an additional 30 minutes, dilute aqueous hydrochloric acid solution (4:1, HZO:HC1) was added until the pH of the reaction mixture reached 2.6. A precipitate of modified amino acids formed and was recovered by decantation. This material was dissolved in 2N sodium hydroxide and dried in vacuo to give a white solid. Yield of crude modified amino acids = 84.6 g, 77W).

MODIFICATION OF PHENYLALANINE METHYL ESTER USING HIPPURYL
CHLORIDE
L-Phenylalanine Methyl Ester Hydrochloride (15 g, 0.084 mole) was dissolved in dimethylformamide (DMF) (100 mL) and to this was added pyridine (30 mL). A solution of hippuryl chloride (16.6 g, 0084 moles in 100 mL DMF) was immediately added to the amino acid ester solution in two portions. The reaction mixture was stirred at room temperature overnight. The reaction mixture was then reduced in vacuo and dissolved in 1N aqueous sodium hydroxide. The solution was heated at 70 C for 3 hours in order to hydrolyze the methyl ester to a free carboxyl group. Thereafter, the solution was acidified to pH 2.25 using dilute aqueous hydrochloric acid solution (1:3 HC1/H2O). A gum-like precipitate formed and this was recovered and dissolved in iN sodium hydroxide. The solution was reduced in vacuo to afford 18.6 g of crude modified amino acid product (Yield 18.6 g). After recrystallization from acetonitrile, pure modified phenylalanine (12 g) was recovered as a white powder. m.p.
223-225 C.

PREPARATION OF DOSING SOLUTIONS:
In a test tube 568 mg of acetyl phenylalanine aldehyde, 132 mg of carbomethoxy phenylalanylleucine and 100 mg acetyl-Phe-Leu-Leu-Arg aldehyde were added to 2.9 ml of 15% ethanol. The solution was stirred and NaOH (1.0 N) was added to ?>arise the pH to 7.2. Water was added to bring the total volume to 4.0 mL. The sample had a carrier concentration of 200 mg/mL. Calcitonin (6 g) was added to the solution. The total calcitonin concentration was 1.5 g/mL.
Following a similar procedure a second solution having 668 mg of acetyl phenylalanine aldehyde and 132 mg of carbomethoxy phenylalanylleucine as the carrier composition and a third solution having as the carrier acetyl phenyl-alanine aldehyde. Each solution had a calcitonin concentration of 1.5 g/mL.

PREPARATION OF MODIFIED AMINO ACID/SALMON CALCITONIN
COMPOSITIONS

(a) Preparation of Modified Amino acid microspheres containing encapsulated Salmon Calcitonin The modified amino acid mixture, prepared in accordance with Example 9, was dissolved at 40 C in distilled water (pH 7.2) at a concentration of 100 mg/ml.
The solution was then filtered with a 0.2 micron filter and the temperature was maintained at 40 C. Salmon calcitonin (Sandoz Corp., Basil, Switzerland) was dissolved in an aqueous solution of citric acid (1.7N) and gelatin (5%-) at a concentration of 150 mg/ml. This solution was then heated to 40 C. The two heated solutions were then mixed 1:1 (v/v). The resulting microsphere suspension was then filtered with glass wool and centrifuged for 50 minutes at 1000 g. The pellet was resuspended with 0.85N citric acid to a volume 5 to 7 fold less than the original volume.
Salmon calcitonin concentration of the resuspended pellet was determined by HPLC. Additional microspheres were made according to the above procedure without salmon calcitonin.
These "empty microspheres" were used to dilute the encapsulated salmon calcitonin microsphere preparation to a final dosing suspension for animal testing.
(b) Preparation of a Soluble Modified Amino acid carrier/Salmon Calcitonin system A soluble amino acid dosing preparation containing salmon calcitonin was prepared by dissolving the modified amino acid material in distilled water (pH 8) to an appropriate concentration. The solution was heated to 40 C
and then filtered with a 0.2 micron filter. Salmon calcitonin, also dissolved in distilled water, was then added to the modified amino acid =solution prior to oral administration.

WO 94/23767 _ 2160693 PCT/US94/04560 ~

.IN VIVO EXPERIMENTS IN RATS
For each sample six fasted rats were anesthetized.
The rats were administered, by oral gavage, one of the 5 calcitonin/carrier dosages prepared in Example 15. The calcitonin concentration in each sample was 1.5 g/ml. Each rat was administered a dosage of two (2) mL/kg each. Blood samples were collected serially from the tail artery. Serum calcium was determined by testing with a DemandTM Calcium Kit 10 (available from Sigma Chemical Company, St. Louis, Missouri, USA). The results of the test are illustrated in Figure 1.

Three samples having 400 mg/kg of acetyl-Leu 15 aldehyde and 10 g/kg of calcitonin, 400 mg/kg of acetyl-Phe aldehyde and 10 g/kg of calcitonin, 200 mg/kg of acetyl-Leu aldehyde, 200 mg/kg of acetyl-Phe aldehyde and 10 g/kg of calcitonin, respectively were prepared. The samples were given to fasted rats as described in Example 16. The 20 results of the test are illustrated graphically in Figure 2.

Two samples having 350 mg/kg of acetyl-Phe aldehyde, 50 mg/kg of carbomethoxy-Phe-Leu-OH and 3 g/kg of 25 calcitonin, 400 mg/kg of acetyl-Phe aldehyde, 50 mg/kg of carbomethoxy-Phe-Leu-OH and 10 g/kg of calcitonin, respectively were prepared. The samples were given to fasted rats as described in Example 16. The results of the test are illustrated in Figure 3.

Three samples having 284 mg/kg of acetyl-Phe aldehyde and 66 mg/kg acetyl-Leu-Leu-Arg aldehyde, 50 mg/kg of carbomethoxy-Phe-Leu-OH and 3 g/kg of calcitonin in propylene glycol, 284 mg/kg of acetyl-Phe aldehyde and 66 mg/kg acetyl-Leu-Leu-Arg aldehyde, 50 mg/kg of carbomethoxy-Phe-Letii-OH and 3 g/kg of calcitonin and 3 jig/kg of calcitonin, in aqueous ethanol, respectively were prepared.

WO 94/23767 ^ 2160693 PCT/US94/04560 The samples were given to fasted rats as described in Example 16. The results of the test are illustrated in Figure 4.

Three samples having 400 mg/kg of 4-(phenylsulfon-amido)-4-phenylbutyric acid and 1.5 g/kg of calcitonin in propylene glycol, 200 mg/kg of 4-(phenylsulfonamido)-4-phenylbutyric acid, 200 mg/kg of acetyl-Phe aldehyde and 1.5 g/kg of calcitonin in aqueous ethanol, respectively were prepared. The samples were given to fasted rats by intraduodenal injection. The results of the test are illustrated in Figure 5.

Samples having 600 mg/kg of acetyl-Phe aldehyde and 10 g/kg of calcitonin in aqueous ethanol, and 3 g/kg of calcitonin, 200 mg/kg of acetyl-Phe aldehyde, 200 mg/kg N-acetyllysinone, 200 mg/kg acetyl-Leu aldehyde and 10 g/kg of calcitonin were prepared. The samples were given to fasted rats as described in Example 16. The results of the test are illustrated in Figure 6.

Three samples having 200 mg/kg of acetyl-Phe aldehyde and 3 g/kg of calcitonin, in aqueous ethanol, dimethyl sulfoxide (DMSO), and olive oil, respectively, were prepared. Additionally a sample of 3 g/kg of calcitonin in DMSO alone was prepared. The samples were given to rats by intraduodenal injection. The results of the test are illustrated in Figure 7.

A sample having 400 mg/kg of cyclohexanoyl-Phe aldehyde and 3 g/kg of calcitonin in aqueous ethanol was prepared. The sample was given to fasted rats as described in Example 16. The results of the test are illustrated in Figure S.

WO 94/23767 216 9 6 9 3 PCT/US94/04560 ~

In vivo evaluation of modified amino acid microspheres containing encapsulated calcitonin and soluble modified amino acid carrier/calcitonin system, prepared as described in Example 16, were evaluated in rats. Rats were gavaged with the oral dosing preparations and blood samples were withdrawn at various time intervals for serum calcium concentration determinations.
Nine rats are divided into three groups as follows:
1. calcitonin microspheres: 10 ug calcitonin/kg body weight by oral gavage (3 rats);
2. calcitonin microspheres: 30 ug calcitonin/kg body weight by oral gavage (3 rats); and 3. soluble modified amino acid/calcitonin system: 30 ug calcitonin/kg body weight by oral gavage (3 rats). The rats were pre-dosed with 0.7 meq of aqueous sodium bicarbonate solution prior to administration of the soluble system.
Oral gavage dosing of rats is performed.
Calcitonin microspheres are prepared immediately prior to dosing and Group 1 rats and Group 2 rats each receive an appropriate dosage of the microsphere suspension. Group 3 rats receives the unencapsulated calcitonin/modified amino acid system. Approximately 0.5 ml of blood is withdrawn from each rat just prior to dosing ("0" time) and 1 h, 2 h and 3 h post-dosing. Serum from the blood samples are stored at -20 C.
The calcium levels of thawed serum taken from group 1-3 rats are analyzed by conventional methods.
Experimental results in rats have demonstrated a significant increase in pharmacological activity (i.e., decreasing serum calcium levels) when calcitonin is orally administered either as a encapsulate in modified amino acid microspheres or a mixture with modified amino acids as compared to basal levels. As shown in Figure 9, soluble modified amino acid solution containing salmon calcitonin demonstrated a significant increase in pharmacological activity (i.e., SUBSTITUTE SHEET (RULE 26) decreasing serum calcium levels) when compared to basal levels after oral administration.

Two samples having 366 mg/kg of acetyl-Phe aldehyde, 33 mg/kg of actinonin and 10 g/kg of calcitonin, 366 mg of acetyl-Phe aldehyde, 33 mg/kg of carbomethoxy-Phe-Leu-OH and 10 g/kg of calcitonin, respectively, were prepared. The samples were given to fasted rats as described in Example 14. The results of the test are illustrated in Figure 10.

Two samples having 400 mg/kg of 4-(phenylsulfonamido)-4-phenylbutyric acid and 3 g/kg of calcitonin, 400 mg/kg of 4-(phenylsulfonamido)-4-phenylbutyric acid and 10 g/kg of calcitonin, respectively were prepared. The samples were given to fasted rats as described in Example 14. The results of the test are illustrated in Figure 11.

Two samples having 400 mg/kg of 3-(phenylsulfonamido)benzoic acid and 10 g/kg of calcitonin, 400 mg/kg of 4-(phenylsulfonamido)hippuric acid and 10 g/kg of calcitonin, respectively were prepared. The samples were given to fasted rats as described in Example 14. The results of the test are illustrated in Figure 12.

Two samples having 400 mg/kg of 4-(phenylsulfonamido)-4-phenylbutyric acid and 10 g/kg of calcitonin, 400 mg/kg of 4-(phenylsulfonamido)benzoic acid . 35 and 10 g/kg of calcitonin, respectively were prepared. The samples were given to fasted rats as described in Example 14. The results of the test are illustrated in Figure 13.
SUBSTITUTE SHEET (RULE 26) Two samples having 400 mg/kg of 4-(phenylsulfona-mido)-4-phenylbutyric acid and 10 g/kg of calcitonin, 400 mg/kg of 4-(phenylsulfonamido)phenylacetic acid and 10 g/kg of calcitonin, respectively were prepared. The samples were given to fasted rats as described in Example 14. The results of the test are illustrated in Figure 14.

In Vivo EVALUATION OF INTERFERON PREPARATIONS IN RATS
Following the procedure described herein samples containing the carriers of the subject invention, in a Trizma hydrochloride buffer solution (Tris-HC1) at a pH of about 7-8, and interferon a2b were prepared. The animals were administered the drug by oral gavage. The delivery was evaluated by using an ELISA assay for human interferon a.

A sample having 800 mg/kg of 4-(phenylsulfona-mido)-4-phenylbutyric acid in a buffered solution and 1000 g/kg of interferon cx2b was prepared. The sample was given to fasted rats as described in Example 14. The results of the test are illustrated in Figure 15.

A sample having 400 mg/kg of 4-(phenylsulfonamido-methyl)benzoic acid in a buffered solution and 1000 g/kg of interferon cx2b was prepared. The sample was given to fasted rats as described in Example 14. The results of the test are illustrated in Figure 16.

A sample having 800 mg/kg of 4-(phenylsulfona-mido)phenylacetic acid in a buffered solution and 1000 g/kg of interferon a2b was prepared. The sample was given to fasted rats as described in Example 14. The results of the test are illustrated in Figure 17.

~~~1~~~7~
~ WO 94/23767 _ PCTIUS94/04560 A sample having 600 mg/kg of 4-(phenylsulfona-mido)hippuric acid in a buffered solution and 1000 g/kg of interferon cx2b was prepared. The sample was given to fasted rats as described in Example 14. The results of the test 5 are illustrated in Figure 18.

In Vivo EVALUATION OF GROWTH HORMONE PREPARATIONS IN RATS
Following the procedure described herein samples containing the carriers of the subject invention and growth 10 hormone were prepared. The animals were administered the drug by oral gavage. The delivery was evaluated by using an ELISA assay for growth hormone.

15 A sample having 1000 mg/kg of 4-(phenylsulfona-mido)-4-phenylbutyric acid and 1 mg/kg of growth hormone was prepared. The sample was given to hypophysectomized rats as.
described in Example 14. The results of the test are illustrated in Figure 19.

A sample having 500 mg/kg of 4-(phenylsulfona-mido)-4-phenylbutyric acid and 1 mg/kg of growth hormone was prepared. In a comparison a group of hypophysectomized rats were given samples of growth hormone without a carrier. The samples were given to hypophysectomized rats as described in Example 14. The results of the test are illustrated in Figure 20.

Two samples having 500 mg/kg of 4-(phenylsulfona-mido)-4-phenylbutyric acid and 6 mg/kg of growth hormone were prepared. The samples were given to normal rats as described in Example 14. The results of the tests are illustrated in Figure 21.

In Vivo EVALUATION OF CROMOGLYCOLATE PREPARATIONS IN RATS
Example 37 Following the procedure described herein samples containing the carriers of the subject invention and disodium cromoglycolate where prepared. The sample, in 0.85N
citric acid and 0.5% acacia, contained 200 mg/kg of 4-(phenylsulfonamido)-4-phenylbutyric acid and 50 mg/kg of disodium cromoglycate. The animals were administered the drug by oral gavage. The delivery was evaluated by using the procedure described by A. Yoshimi in Pharmacobio-Dyn., 15, pages 681-686, (1992). The results of the tests are illustrated in Figure 22.

As clearly illustrated by the data in the Examples and Figures the use of compositions of the subject invention show significant advantages for the delivery of biologically active agents.

Many variations of the present invention will suggest themselves to those skilled in the art in light of the above detailed disclosure. For example, poly(amino acids) which are formed by a bond other than an amide bound, e.g., an ester or an anhydride linkage, may be derivatized and modified for use as carriers in accordance with the present invention. All such modifications are within the full intended scope of the appended claims.

Claims (11)

WHAT IS CLAIMED IS:
1. An oral composition comprising:

(A) at least one biologically-active agent selected from the group consisting of human growth hormone, bovine growth hormone, growth hormone-releasing hormone, an interferon, interleukin-I, interleukin-II, insulin, heparin, low molecular weight heparin, calcitonin, erythropoietin, atrial naturetic factor, an antigen, a monoclonal antibody, somatostatin, adrenocorticotropin, gonadotropin releasing hormone, oxytocin, vasopressin, cromolyn sodium, vancomycin, desferrioxamine and any combination thereof; and (B) a compound having the formula:
Ar-Y-(R1)n-OH
wherein:
Ar is a phenyl or naphthyl optionally substituted with C1 to C6 alkyl, C1 to C6 alkenyl, C1 to C6 alkoxy, hydroxy, thio, or CO2R6 wherein R6 is hydrogen, C1 to C6 alkyl or C1 to C6 alkenyl;

; wherein:

R3 is C1 to C24 alkyl, C1 to C24 alkenyl, phenyl, naphthyl, (C1 to C10 alkyl) phenyl, (C1 to C10 alkenyl) phenyl, (C1 to C10 alkyl) naphthyl, (C1 to alkenyl) naphthyl, phenyl (C1 to C10 alkyl), phenyl (C1 to C10 alkenyl), naphthyl (C1 to C10 alkyl), or naphthyl (C1 to C10 alkenyl);
R3 is unsubstituted or substituted with C1 to C4 alkyl, C1 to C4 alkenyl, C1 to C4 alkoxy, -OH, -SH, -CO2R5 , cycloalkyl, cycloalkenyl, heterocyclic, aryl, alkaryl, heteroaryl, heteroalkaryl or any combination thereof;
R5 is hydrogen, C1 to C4 alkyl or C1 to C4 alkenyl;

R3 is uninterrupted or interrupted by oxygen, nitrogen, sulfur or any combination thereof; and R4 is hydrogen, C1 to C4 alkyl or C1 to C4 alkenyl; and n is an integer from 1 to 5.
2. The composition according to claim 1, wherein:
Ar is a phenyl optionally substituted with C1 to C6 alkyl, C1 to C6 alkenyl, C1 to C6 alkoxy, hydroxy, thio, or CO2R6 wherein R6 is hydrogen, C1 to C6 alkyl or C1 to C6 alkenyl;

Y is , R3 is C1 to C24 alkyl, (C1 to C10 alkyl) phenyl or phenyl (C1 to C10 alkyl);
and n is equal to 1.
3. The composition according to claim 1 or 2, wherein said biologically-active agent comprises an interferon, insulin, growth hormone, heparin, calcitonin, cromolyn sodium.
4. The composition according to claim 1 or 2, wherein said biologically-active agent is calcitonin.
5. The composition according to claim 1 or 2, wherein said biologically-active agent is growth hormone.
6. The composition according to claim 1 or 2, wherein said biologically-active agent is interferon.
7. The composition according to claim 1 or 2, wherein said biologically-active agent is cromolyn sodium.
8. A dosage unit form comprising:

(A) an oral composition as defined in any one of claims 1 to 7;
and (B) (a) an excipient, (b) a diluent, (c) a disintegrant, (d) a lubricant, (e) a plasticizer, (f) a colorant, (g) a dosing vehicle, or (h) any combination thereof.
9. The dosage unit form according to claim 10 comprising a tablet, a capsule, or a liquid.
10. Use of an oral composition as defined in any one of claims 1 to 7 for the preparation of a medicament for oral administration to an animal.
11. A method for preparing the oral composition according to claim 1, said method comprising mixing:

(A) at least one biologically-active agent selected from the group consisting of human growth hormone, bovine growth hormone, growth hormone-releasing hormone, an interferon, interleukin-I, interleukin-II, insulin, heparin, low molecular weight heparin, calcitonin, erythropoietin, atrial naturetic factor, an antigen, a monoclonal antibody, somatostatin, adrenocorticotropin, gonadotropin releasing hormone, oxytocin, vasopressin, cromolyn sodium, vancomycin, desferrioxamine and any combination thereof; and (B) a compound having the formula:
Ar-Y-(R1)n-OH
wherein:

Ar is a phenyl or naphthyl optionally substituted with C1 to C6 alkyl, C1 to C6 alkenyl, C1 to C6 alkoxy, hydroxy, thio, or CO2R6 wherein R6 is hydrogen, C1 to C6 alkyl or C1 to C6 alkenyl;

; wherein:
R3 is C1 to C24 alkyl, C1 to C24 alkenyl, phenyl, naphthyl, (C1 to C10 alkyl) phenyl, (C1 to C10 alkenyl) phenyl, (C1 to C10 alkyl) naphthyl, (C1 to alkenyl) naphthyl, phenyl (C1 to C10 alkyl), phenyl (C1 to C10 alkenyl), naphthyl (C1 to C10 alkyl), or naphthyl (C1 to C10 alkenyl);
R3 is unsubstituted or substituted with C1 to C4 alkyl, C1 to C4 alkenyl, C1 to C4 alkoxy, -OH, -SH, -CO2R5 , cycloalkyl, cycloalkenyl, heterocyclic, aryl, alkaryl, heteroaryl, heteroalkaryl or any combination thereof;
R5 is hydrogen, C1 to C4 alkyl or C1 to C4 alkenyl;
R3 is uninterrupted or interrupted by oxygen, nitrogen, sulfur or any combination thereof; and R4 is hydrogen, C1 to C4 alkyl or C1 to C4 alkenyl; and n is and integer from 1 to 5.
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Families Citing this family (104)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5578323A (en) 1992-06-15 1996-11-26 Emisphere Technologies, Inc. Proteinoid carriers and methods for preparation and use thereof
US6344213B1 (en) 1996-03-29 2002-02-05 Emisphere Technologies, Inc. Compounds and compositions for delivering active agents
US5447728A (en) * 1992-06-15 1995-09-05 Emisphere Technologies, Inc. Desferrioxamine oral delivery system
US6331318B1 (en) 1994-09-30 2001-12-18 Emisphere Technologies Inc. Carbon-substituted diketopiperazine delivery systems
US6916489B2 (en) * 1992-06-15 2005-07-12 Emisphere Technologies, Inc. Active agent transport systems
US5981719A (en) 1993-03-09 1999-11-09 Epic Therapeutics, Inc. Macromolecular microparticles and methods of production and use
US6090925A (en) 1993-03-09 2000-07-18 Epic Therapeutics, Inc. Macromolecular microparticles and methods of production and use
US5643957A (en) * 1993-04-22 1997-07-01 Emisphere Technologies, Inc. Compounds and compositions for delivering active agents
US20010003001A1 (en) 1993-04-22 2001-06-07 Emisphere Technologies, Inc. Compounds and compositions for delivering active agents
US6461643B2 (en) 1993-04-22 2002-10-08 Emisphere Technologies, Inc. Oral drug delivery compositions and methods
EP0696208B1 (en) * 1993-04-22 2001-08-22 Emisphere Technologies, Inc. Oral drug delivery compositions
US5709861A (en) * 1993-04-22 1998-01-20 Emisphere Technologies, Inc. Compositions for the delivery of antigens
EP1792624A1 (en) * 1995-03-31 2007-06-06 Emisphere Technologies, Inc. Modified amino acids and compositions comprising them for delivering active agents
HU229049B1 (en) * 1995-03-31 2013-07-29 Emisphere Tech Inc Compound and compositions for delivering active ingredients
US6001347A (en) 1995-03-31 1999-12-14 Emisphere Technologies, Inc. Compounds and compositions for delivering active agents
US5650386A (en) * 1995-03-31 1997-07-22 Emisphere Technologies, Inc. Compositions for oral delivery of active agents
KR100489667B1 (en) * 1995-03-31 2006-01-12 에미스페어 테크놀로지스, 인코포레이티드 Compounds and compositions for delivering active agents
US5667806A (en) * 1995-06-07 1997-09-16 Emisphere Technologies, Inc. Spray drying method and apparatus
CA2243643A1 (en) 1996-11-18 1998-05-28 Susan Haas Methods and compositions for inducing oral tolerance in mammals
US6358504B1 (en) 1997-02-07 2002-03-19 Emisphere Technologies, Inc. Compounds and compositions for delivering active agents
US6313088B1 (en) 1997-02-07 2001-11-06 Emisphere Technologies, Inc. 8-[(2-hydroxy-4-methoxy benzoyl) amino]-octanoic acid compositions for delivering active agents
JPH11279040A (en) 1998-03-27 1999-10-12 Kao Corp Composition for external use for skin
EP1100771B1 (en) 1998-07-27 2005-05-11 Emisphere Technologies, Inc. Compounds and compositions for delivering active agents
US6440929B1 (en) 1998-07-27 2002-08-27 Emisphere Technologies, Inc. Pulmonary delivery of active agents
US6991798B1 (en) 1998-08-07 2006-01-31 Emisphere Technologies, Inc. Compounds and compositions for delivering active agents
KR100659753B1 (en) * 1998-08-07 2006-12-20 에미스페어 테크놀로지스, 인코포레이티드 Compounds and compositions for delivering active agents
NZ512581A (en) 1999-01-08 2002-12-20 Univ Virginia Commonwealth Polymeric delivery agents comprising a polymer conjugated to a modified amino acid and derivatives thereof
US7084279B1 (en) 1999-02-11 2006-08-01 Emisphere Technologies Inc. Oxadiazole compounds and compositions for delivering active agents
MXPA01008611A (en) 1999-02-26 2003-06-24 Emisphere Tech Inc Compounds and compositions for delivering active agents.
EP1175390B1 (en) * 1999-04-05 2005-02-02 Emisphere Technologies, Inc. Disodium salts, monohydrates, and ethanol solvates
ATE296545T1 (en) * 1999-04-27 2005-06-15 Aionix Invest Ltd SUPPLEMENT FOR RESTORING GROWTH HORMONE LEVELS
US9006175B2 (en) 1999-06-29 2015-04-14 Mannkind Corporation Potentiation of glucose elimination
US6375948B1 (en) * 1999-07-12 2002-04-23 Kao Corporation Treating method for suppressing hair growth
US7279597B1 (en) 1999-11-05 2007-10-09 Emisphere Technologies, Inc. Phenyl amine carboxylic acid compounds and compositions for delivering active agents
US7129274B1 (en) 1999-11-05 2006-10-31 Emisphere Technologies Inc. Phenoxy carboxylic acid compounds and compositions for delivering active agents
EP1237872B1 (en) 1999-12-16 2008-02-27 Emisphere Technologies, Inc. Compounds and compositions for delivering active agents
AU2001236457A1 (en) 2000-01-13 2001-07-24 Emisphere Technologies, Inc. Compounds and compositions for delivering active agents
US7227033B2 (en) 2002-01-09 2007-06-05 Emisphere Technologies, Inc. Polymorphs of sodium 4-[(4-chloro-2-hydroxybenzoyl) amino] butanoate
IL153297A0 (en) 2000-06-29 2003-07-06 Emisphere Tech Inc Compounds and compositions for delivering active agents
CN1313441C (en) 2000-09-06 2007-05-02 艾米斯菲尔技术有限公司 Cyanophenoxy carboxylic acid compounds and compositions for delivering active agents
AU7338801A (en) * 2000-09-08 2002-03-22 Gryphon Sciences "pseudo"-native chemical ligation
AU2002245580B2 (en) 2001-03-01 2006-11-09 Emisphere Technologies, Inc. Compounds and compositions for delivering active agents
US20030225300A1 (en) * 2001-04-19 2003-12-04 Emisphere Technologies Inc. Compounds and compositions for delivering active agents
US7008650B2 (en) * 2001-08-09 2006-03-07 Lam Paul Y S Compositions for the treatment of acquired immunodeficiency disease
CA2466863A1 (en) * 2001-11-29 2003-06-05 Emisphere Technologies, Inc. Oral pharmaceutical compositions comprising cromolyn sodium and an acylated amino acid
JP4417113B2 (en) 2002-02-20 2010-02-17 エミスフェアー・テクノロジーズ・インク Administration method of GLP-1 molecule
EP1894591B1 (en) 2002-03-20 2013-06-26 MannKind Corporation Cartridge for an inhalation apparatus
JP2006528982A (en) * 2003-05-14 2006-12-28 エミスフェアー・テクノロジーズ・インク Composition for delivery of peptide YY and PYY agonists
FR2855521B1 (en) * 2003-05-28 2005-08-05 Flamel Tech Sa POLYAMINOACIDES FUNCTIONALIZED BY AT LEAST ONE YDROPHOBIC GROUP AND THEIR PARTICULARLY THERAPEUTIC APPLICATIONS.
US20060286129A1 (en) * 2003-12-19 2006-12-21 Emisphere Technologies, Inc. Oral GLP-1 formulations
JP2007532684A (en) * 2004-04-16 2007-11-15 エミスフェアー・テクノロジーズ・インク 8- (2-Hydroxyphenoxy) octyldiethanolamine and its salts for delivery of active agents
US8273794B2 (en) 2004-05-14 2012-09-25 Emisphere Technologies, Inc. Compounds and compositions for delivering active agents
JP4995080B2 (en) * 2004-05-14 2012-08-08 エミスフェアー・テクノロジーズ・インク Aryl ketone compounds and compositions for delivering active agents
EP1750718B1 (en) 2004-05-19 2014-08-13 Emisphere Technologies, Inc. Acyclovir formulations
US9498487B2 (en) 2004-05-19 2016-11-22 Emisphere Technologies, Inc. Topical cromolyn formulations
AU2005271526B2 (en) 2004-08-03 2011-12-08 Emisphere Technologies, Inc. Antidiabetic oral insulin-biguanide combination
WO2006124047A2 (en) 2004-08-13 2006-11-23 Emisphere Technologies, Inc. Pharmaceutical formulations containing microparticles or nanoparticles of a delivery agent
US7709639B2 (en) 2004-08-20 2010-05-04 Mannkind Corporation Catalysis of diketopiperazine synthesis
EP1791542B1 (en) 2004-08-23 2015-05-20 Mannkind Corporation Diketopiperazine salts for drug delivery
BRPI0517574A (en) 2004-12-29 2008-10-14 Emisphere Tech Inc pharmaceutical formulation, method for treating or preventing hypercalcemia and method for preparing pharmaceutical formulation
US8975227B2 (en) 2005-07-15 2015-03-10 Emisphere Technologies, Inc. Intraoral dosage forms of glucagon
US8541362B2 (en) * 2005-08-19 2013-09-24 Emisphere Technologies, Inc. Cyclopropyl compounds and compositions for delivering active agents
US20070049557A1 (en) * 2005-08-24 2007-03-01 Hashim Ahmed Solid pharmaceutical dosage forms comprising bisphosphonates and modified amino acid carriers
ES2559677T3 (en) 2005-09-14 2016-02-15 Mannkind Corporation Drug formulation method based on increasing the affinity of the active ingredients for microcrystalline particle surfaces
KR20080096809A (en) 2006-02-22 2008-11-03 맨카인드 코포레이션 A method for improving the pharmaceutic properties of microparticles comprising diketopiperazine and an active agent
WO2007121318A2 (en) 2006-04-12 2007-10-25 Emisphere Technologies, Inc. Formulations for delivering insulin
WO2007121471A2 (en) * 2006-04-18 2007-10-25 Emisphere Technologies, Inc. Dialkyl ether delivery agents
JP5475443B2 (en) * 2006-06-28 2014-04-16 エミスフェアー・テクノロジーズ・インク Gallium nitrate preparation
WO2008014430A1 (en) 2006-07-27 2008-01-31 Emisphere Technologies, Inc. Arylsulfanyl compounds and compositions for delivering active agents
JP5564255B2 (en) 2006-08-18 2014-07-30 エミスフェアー・テクノロジーズ・インク Synthesis of propylphenoxy ether and use as a delivery agent
US8895777B2 (en) * 2006-08-31 2014-11-25 Emisphere Technologies Inc Compounds and compositions for delivering active agents
CN105233294A (en) * 2007-02-08 2016-01-13 爱密斯菲尔科技公司 Phenylalkylcarboxylic acid delivery agent
CA2678575C (en) * 2007-02-16 2013-10-08 Emisphere Technologies, Inc. Compounds having a cyclic moiety and compositions for delivering active agents
PL2134351T3 (en) 2007-03-13 2017-10-31 Jds Therapeutics Llc Methods and compositions for the sustained release of chromium
CA2680737C (en) * 2007-03-21 2015-12-15 Emisphere Technologies, Inc. Allyloxy and alkyloxy benzoic acid delivery agents
US20080260823A1 (en) * 2007-04-20 2008-10-23 Sciele Pharma, Inc. Orally disintegrating tablet comprising glycopyrrolate for treating sialorrhea
WO2009002867A2 (en) 2007-06-26 2008-12-31 Nutrition 21, Inc. Multiple unit dosage form having a therapeutic agents in combination with a nutritional supplement
WO2009033667A2 (en) * 2007-09-11 2009-03-19 Mondobiotech Laboratories Ag Use of a peptide as a therapeutic agent
CA3086027C (en) 2008-06-13 2022-05-17 Mannkind Corporation A dry powder inhaler and system for drug delivery
US8485180B2 (en) 2008-06-13 2013-07-16 Mannkind Corporation Dry powder drug delivery system
DK2300083T3 (en) 2008-06-20 2013-07-22 Mannkind Corp INTERACTIVE DEVICE AND PROCEDURE FOR REAL-TIME PROFILING INHALATION TESTS
TWI532497B (en) 2008-08-11 2016-05-11 曼凱公司 Use of ultrarapid acting insulin
US8314106B2 (en) 2008-12-29 2012-11-20 Mannkind Corporation Substituted diketopiperazine analogs for use as drug delivery agents
EP2405963B1 (en) 2009-03-11 2013-11-06 MannKind Corporation Apparatus, system and method for measuring resistance of an inhaler
EP2236617A1 (en) * 2009-03-31 2010-10-06 Leukocare Ag Methods of terminal sterilization of biofunctional compositions
BRPI1013154B1 (en) 2009-06-12 2020-04-07 Mannkind Corp MICROPARTICLES OF DICETOPIPERAZINE WITH SPECIFIC SURFACE AREAS DEFINED, DRY POWDER UNDERSTANDING THE REFERRED MICROPARTICLES, METHOD FOR FORMATION OF THE REFERENCESMICROPARTICLES AND THE FORMATION OF MICROPARTYSTEMS
JP5985985B2 (en) 2009-08-03 2016-09-06 エミスフィアー テクノロジーズ インコーポレイテッドEmisphere Technologies,Inc. Rapid-acting naproxen composition with reduced gastrointestinal effects
WO2011056889A1 (en) 2009-11-03 2011-05-12 Mannkind Corporation An apparatus and method for simulating inhalation efforts
RU2531455C2 (en) 2010-06-21 2014-10-20 Маннкайнд Корпорейшн Systems and methods for dry powder drugs delivery
EP4070801A1 (en) 2011-03-01 2022-10-12 Nutrition 21, LLC Compositions of insulin and chromium for the treatment and prevention of diabetes, hypoglycemia and related disorders
MX353285B (en) 2011-04-01 2018-01-05 Mannkind Corp Blister package for pharmaceutical cartridges.
WO2012174472A1 (en) 2011-06-17 2012-12-20 Mannkind Corporation High capacity diketopiperazine microparticles
KR20140095483A (en) 2011-10-24 2014-08-01 맨카인드 코포레이션 Methods and compositions for treating pain
RU2650035C2 (en) 2012-07-12 2018-04-06 Маннкайнд Корпорейшн Dry powder drug delivery systems and methods
EP2888224B1 (en) 2012-08-23 2017-11-15 Emisphere Technologies, Inc. Phenoxy alkyl diethanolamine and diisopropanolamine compounds for delivering active agents
SG10202011046RA (en) 2012-09-21 2020-12-30 Intensity Therapeutics Inc Method of treating cancer
WO2014066856A1 (en) 2012-10-26 2014-05-01 Mannkind Corporation Inhalable influenza vaccine compositions and methods
CN105102436B (en) 2013-03-15 2018-06-12 曼金德公司 Crystallite diketopiperazine composition and method
KR102321339B1 (en) 2013-07-18 2021-11-02 맨카인드 코포레이션 Heat-stable dry powder pharmaceutical compositions and methods
CA2920488C (en) 2013-08-05 2022-04-26 Mannkind Corporation Insufflation apparatus and methods
WO2015148905A1 (en) 2014-03-28 2015-10-01 Mannkind Corporation Use of ultrarapid acting insulin
US10561806B2 (en) 2014-10-02 2020-02-18 Mannkind Corporation Mouthpiece cover for an inhaler
MX2017008913A (en) 2015-01-07 2018-04-30 Trigemina Inc Magnesium-containing oxytocin formulations and methods of use.
WO2017139337A1 (en) 2016-02-11 2017-08-17 Nutrition 21, Llc Chromium containing compositions for improving health and fitness

Family Cites Families (136)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US24899A (en) * 1859-07-26 Edwabd f
US2671451A (en) * 1952-06-16 1954-03-09 Stephen J Bolger Remedial pill
BE530010A (en) 1953-06-30
US2868740A (en) * 1954-03-25 1959-01-13 Swift & Co Method of copolymerizing acrylic or methacrylic acid with proteinaceous material and product obtained
US2862918A (en) * 1956-03-12 1958-12-02 Glidden Co Acylated, isolated, partially-hydrolyzed, soya protein and process
NL108169C (en) * 1957-01-30
US3057344A (en) * 1957-05-21 1962-10-09 Abella Carlos Alberto Capsule for the study of the digestive tract and method of using the same
US3016308A (en) * 1957-08-06 1962-01-09 Moore Business Forms Inc Recording paper coated with microscopic capsules of coloring material, capsules and method of making
US3052655A (en) * 1958-08-01 1962-09-04 Sidney W Fox Thermal polymerization of amino acid mixtures containing aspartic acid or a thermal precursor of aspartic acid
US3076790A (en) * 1958-08-01 1963-02-05 Sidney W Fox Method of making copolymers of amino acids containing glutamic acid
FR1468601A (en) 1958-12-22 1967-02-10 Ncr Co Process for forming protective coatings for solid and liquid particles
FR1351358A (en) 1958-12-22 1964-02-07 Ncr Co Process for forming impermeable coatings for particulate matter by liquid phase separation
GB929401A (en) 1958-12-22 1963-06-19 Upjohn Co Encapsulated emulsions and processes for their preparation
NL246985A (en) * 1958-12-31
US3170802A (en) * 1960-12-14 1965-02-23 Zh Noda Sangyo Kagaku Kenkyush Method for treatment of soybean proteins
GB1075952A (en) 1962-12-31 1967-07-19 Gelatine And Glue Res Ass Microscopic capsules and methods of making them
US3748277A (en) * 1965-10-14 1973-07-24 Ncr Co Process of forming minute capsules
US3474777A (en) * 1966-02-10 1969-10-28 Amp Inc Method of administering therapeutic agents
US3576758A (en) * 1966-10-17 1971-04-27 Ncr Co Treatment of polypeptide-containing hydrophilic polymeric capsule wall material with uranium and vanadium compounds
FR7981M (en) * 1967-10-21 1970-06-08
US3491093A (en) * 1967-11-29 1970-01-20 Endo Lab Derivatives of 5 aminomethyl-4,5,6,7-tetrahydro-4-oxoindoles
US3565559A (en) * 1968-03-11 1971-02-23 Sumitomo Chemical Co Process for making microcapsules
US3574832A (en) * 1968-05-29 1971-04-13 American Cyanamid Co Therapeutic heparin-surfactant compositions
US3567650A (en) * 1969-02-14 1971-03-02 Ncr Co Method of making microscopic capsules
US3937668A (en) * 1970-07-15 1976-02-10 Ilse Zolle Method for incorporating substances into protein microspheres
US3725113A (en) * 1970-12-17 1973-04-03 Research Corp Blood compatible microencapsulated detoxicants and method for making
US3822348A (en) * 1970-12-28 1974-07-02 Toyo Jozo Kk Hormone-like substance having serum calcium reducing property
US3962416A (en) * 1971-01-25 1976-06-08 Sol Katzen Preserved nutrients and products
IL36670A (en) 1971-04-21 1974-09-10 Sela M Therapeutic basic copolymers of amino acids
US3794561A (en) * 1971-09-30 1974-02-26 Sasaki T Biologically active peptide and method of preparing the same
US3816404A (en) * 1971-12-08 1974-06-11 Texaco Inc Preparation of caprolactam
US3933873A (en) * 1971-12-08 1976-01-20 Texaco Inc. Preparation of omega-aminoalkanoic acids
US3795739A (en) * 1972-02-14 1974-03-05 Hoffmann La Roche Treatment of parkinson disease
JPS5210427B2 (en) * 1972-07-19 1977-03-24
US4450150A (en) * 1973-05-17 1984-05-22 Arthur D. Little, Inc. Biodegradable, implantable drug delivery depots, and method for preparing and using the same
CA1045977A (en) 1973-05-17 1979-01-09 Arthur D. Little Biodegradable, implantable drug delivery device, and process for preparing and using the same
US4351337A (en) * 1973-05-17 1982-09-28 Arthur D. Little, Inc. Biodegradable, implantable drug delivery device, and process for preparing and using the same
DE2343037A1 (en) 1973-08-25 1975-03-06 Hoechst Ag MEDICINAL PRODUCTS WITH ANTIDEPRESSIVE EFFECT
US3939253A (en) * 1973-11-02 1976-02-17 Interx Research Corporation Novel, transient pro-drug forms of l-dopa useful in the treatment of parkinson's disease
GB1459488A (en) * 1974-03-19 1976-12-22 Wyeth John & Brother Ltd Piperazinedione derivatives
US4061466A (en) * 1974-10-16 1977-12-06 Ingvar Gosta Holger Sjoholm Biologically active composition and the use thereof
US4183849A (en) * 1975-01-15 1980-01-15 Nordisk Insulinlaboratorium Therapeutic insulin preparation and a process for the production of a stable insulin preparation with protracted effect
US4048268A (en) * 1975-02-19 1977-09-13 Eli Lilly And Company Stabilization method
US4035507A (en) * 1975-04-17 1977-07-12 Interx Research Corporation Novel, transient pro-drug forms of L-DOPA to treat Parkinson's disease
CA1077842A (en) 1975-10-09 1980-05-20 Minnesota Mining And Manufacturing Company Albumin medicament carrier system
US4405598A (en) * 1976-01-30 1983-09-20 Fisons, Limited Composition for treating asthma
US4117801A (en) * 1976-06-10 1978-10-03 Eastman Kodak Company Apparatus for spray coating discrete particles
US4357259A (en) * 1977-08-01 1982-11-02 Northwestern University Method of incorporating water-soluble heat-sensitive therapeutic agents in albumin microspheres
US4217370A (en) * 1977-08-25 1980-08-12 Blue Wing Corporation Lipid-containing feed supplements and foodstuffs
US4199561A (en) * 1979-02-26 1980-04-22 The Dow Chemical Company Coated nutrients and medicaments for veterinary use
US4352883A (en) * 1979-03-28 1982-10-05 Damon Corporation Encapsulation of biological material
US4345588A (en) * 1979-04-23 1982-08-24 Northwestern University Method of delivering a therapeutic agent to a target capillary bed
US4272506A (en) * 1979-08-31 1981-06-09 Syva Company Purification of reagents by disulfide immobilization
HU181009B (en) * 1980-01-18 1983-05-30 Richter Gedeon Vegyeszet Process for preparing angiotensin-ii analogues with antagonictic activity containing in position 1 sarcosyl,hydroxyacetyl or l-alpha-aminoxy-propionyl group and in positiona 8 esteric group
NZ196349A (en) * 1980-03-07 1984-08-24 Interx Research Corp Enhancement of absorption rate of orally administered polar bioactive agents
IT1148784B (en) * 1980-04-09 1986-12-03 Eurand Spa PROCEDURE FOR THE PREPARATION OF MICRO CAPSULES IN A LIQUID VEHICLE
DE3016170A1 (en) * 1980-04-26 1981-10-29 Bayer Ag, 5090 Leverkusen MICROCAPSULES WITH A DEFINED OPENING TEMPERATURE, METHOD FOR THE PRODUCTION AND USE THEREOF
US4289759A (en) * 1980-06-23 1981-09-15 Ortho Pharmaceutical Corporation Immunoregulatory diketopiperazine compounds
US4348384A (en) * 1980-10-17 1982-09-07 Dainippon Pharmaceutical Co., Ltd. Pharmaceutical composition for oral administration containing coagulation factor VIII or IX
US4442090A (en) * 1980-11-09 1984-04-10 Kyoto Yakuhin Kogyo Kabushiki Kaisha Absorption-promoting compounds, compositions thereof with pharmaceuticals and/or bases for rectal administration and method of use
US4900730A (en) * 1981-01-14 1990-02-13 Toyo Jozo Co., Ltd. Preparation which promotes the absorption of peptides
GB2092136B (en) * 1981-01-17 1985-06-05 Mitsui Toatsu Chemicals Production of n-substituted amide compounds
US4483807A (en) 1981-01-27 1984-11-20 Japan Atomic Energy Research Institute Process for producing a slow release composite
FR2509175B1 (en) * 1981-03-06 1987-01-16 Toyo Jozo Kk THERAPEUTIC PREPARATION HAVING EXCELLENT ABSORPTION PROPERTIES
JPS57146721A (en) * 1981-03-06 1982-09-10 Toyo Jozo Co Ltd Improver for absorption
JPS58140026A (en) * 1982-01-14 1983-08-19 Toyo Jozo Co Ltd Pharmaceutical having improved absorbability
NZ201010A (en) 1981-06-19 1986-02-21 Ciba Geigy Ag The treatment of inflammation diseases using desferrioxamine
US4446138A (en) * 1982-02-10 1984-05-01 Pack Howard M Method and composition for reducing weight
CA1241646A (en) * 1982-02-22 1988-09-06 Adolfo J. De Bold Atrial natriuretic factor
US4518433A (en) * 1982-11-08 1985-05-21 Fmc Corporation Enteric coating for pharmaceutical dosage forms
US4473620A (en) * 1982-12-23 1984-09-25 Eastman Kodak Company Encapsulated butylated hydroxyanisole
US4886663A (en) * 1983-01-03 1989-12-12 Scripps Clinic And Research Foundation Synthetic heat-stable enterotoxin polypeptide of Escherichia coli and multimers thereof
JPS59163313A (en) * 1983-03-09 1984-09-14 Teijin Ltd Peptide hormone composition for nasal administration
CA1196862A (en) * 1983-06-01 1985-11-19 Anthony M.F. Sun Microencapsulation of living tissue and cells
CA1196863A (en) * 1983-06-08 1985-11-19 Mattheus F.A. Goosen Slow release injectable insulin composition
US4462839A (en) * 1983-06-16 1984-07-31 Fmc Corporation Enteric coating for pharmaceutical dosage forms
US4608278A (en) * 1983-06-22 1986-08-26 The Ohio State University Research Foundation Small particule formation and encapsulation
US4671954A (en) * 1983-12-13 1987-06-09 University Of Florida Microspheres for incorporation of therapeutic substances and methods of preparation thereof
US4590265A (en) * 1984-02-17 1986-05-20 Eastman Kodak Company Carboxylated cellulose ester and manufacture thereof
JPS60176549A (en) * 1984-02-22 1985-09-10 Nisshin Oil Mills Ltd:The Preparation of protein hydrolyzate
US4703042A (en) * 1984-05-21 1987-10-27 Bodor Nicholas S Orally active heparin salts containing multivalent cationic units
FR2565102B1 (en) * 1984-06-05 1987-03-20 Paris Sud Universite BIODEGRADABLE MICROCAPSULES BASED ON SERUMALBUMIN, THEIR PREPARATION AND THEIR APPLICATION TO THE IN SITU RELEASE OF MEDICUMENTS
US4757066A (en) * 1984-10-15 1988-07-12 Sankyo Company Limited Composition containing a penem or carbapenem antibiotic and the use of the same
IT1177384B (en) * 1984-12-12 1987-08-26 Boeehringer Biochemia Robin Sp DIETARY GRANULAR PRODUCTS BASED ON AMINO ACIDS AND PROCEDURE FOR THEIR PREPARATION
US4708952A (en) * 1985-02-06 1987-11-24 Aida Salatinjants Method of treatment of the infectious and viral diseases by one time interference
CS254355B1 (en) * 1985-04-10 1988-01-15 Vladimir Saudek Soluble and biodegradatable copolymeres activated for bond of biologicaly active substances
US4897444A (en) * 1985-05-31 1990-01-30 The Research Foundation Of The State University Of New York Immobilized fluorogenic substrates for enzymes; and processes for their preparation
US4757024A (en) * 1985-05-31 1988-07-12 Biostar Medical Products, Inc. Immune complex detection method and article using immunologically non-specific immunoglobulins
US4789734A (en) * 1985-08-06 1988-12-06 La Jolla Cancer Research Foundation Vitronectin specific cell receptor derived from mammalian mesenchymal tissue
IT1214629B (en) * 1985-08-29 1990-01-18 Formenti Farmaceutici Spa MICRO-ENCAPSULATION PROCEDURE OF A MEDICATION, MEDICATION SO PREPARED, AND PHARMACEUTICAL COMPOSITIONS THAT INCLUDE IT
EP0225130B1 (en) * 1985-11-22 1991-10-30 Takeda Chemical Industries, Ltd. Liposome composition
IT1188550B (en) * 1986-02-07 1988-01-14 Sclavo Spa SYNTHETIC PEPTIDE WITH INTERLEUKINA 1 HUMAN ACTIVITY
US4919939A (en) * 1986-04-29 1990-04-24 Pharmetrix Corporation Periodontal disease treatment system
US4837381A (en) * 1986-08-11 1989-06-06 American Cyanamid Company Compositions for parenteral administration and their use
NZ221411A (en) 1986-08-11 1989-10-27 Innovata Biomed Ltd Pharmaceutical compositions containing microcapsules and a surfactant
US4925673A (en) * 1986-08-18 1990-05-15 Clinical Technologies Associates, Inc. Delivery systems for pharmacological agents encapsulated with proteinoids
DE3700128A1 (en) * 1987-01-03 1988-07-14 Hoechst Ag BIODEGRADABLE POLY- (HYDROXYALKYL) - AMINODICARBONIC ACID DERIVATIVES, METHOD FOR THE PRODUCTION AND USE THEREOF FOR DEPOT PREPARATIONS WITH CONTROLLED ACTIVE SUBSTANCE DELIVERY
US5077278A (en) * 1987-01-23 1991-12-31 Pfizer Inc. Non-natural demethylavermectins compositions and method of use
US5069936A (en) * 1987-06-25 1991-12-03 Yen Richard C K Manufacturing protein microspheres
MX12394A (en) 1987-07-23 1993-12-01 Ciba Geigy Ag PROCEDURE FOR OBTAINING POLYETHYLENE GLYCOL CARBAMATES.
US4895725A (en) * 1987-08-24 1990-01-23 Clinical Technologies Associates, Inc. Microencapsulation of fish oil
US5067961A (en) * 1988-02-18 1991-11-26 Autogenesis Technologies, Inc. Non-biodegradable two phase corneal implant and method for preparing same
JP2670680B2 (en) * 1988-02-24 1997-10-29 株式会社ビーエムジー Polylactic acid microspheres containing physiologically active substance and method for producing the same
US5055300A (en) * 1988-06-17 1991-10-08 Basic Bio Systems, Inc. Time release protein
FR2636238B1 (en) * 1988-09-14 1994-01-21 Morelle Jean NEW ANTISUDORAL COMPOSITIONS
GB8822857D0 (en) 1988-09-29 1988-11-02 Patralan Ltd Pharmaceutical formulations
GB8823731D0 (en) 1988-10-10 1988-11-16 Smith Kline French Lab Biologically active compounds
US5039481A (en) * 1988-12-16 1991-08-13 Clean Air, Inc. Aliphatic polycarboxylic acids as air purification compositions
US4976968A (en) * 1989-02-24 1990-12-11 Clinical Technologies Associates, Inc. Anhydrous delivery systems for pharmacological agents
US4983402A (en) * 1989-02-24 1991-01-08 Clinical Technologies Associates, Inc. Orally administerable ANF
CA2012306A1 (en) 1989-03-28 1990-09-28 Werner Neidhart Amino acid derivatives
US5122367A (en) * 1989-03-31 1992-06-16 Massachusetts Institute Of Technology Polyanhydride bioerodible controlled release implants for administration of stabilized growth hormone
US4963364A (en) * 1989-04-10 1990-10-16 Fox Sidney W Microencapsulated antitumor agent
US5100918A (en) * 1989-05-25 1992-03-31 Sterling Drug, Inc. Prevention or treatment of sunburn using the S(+) isomer of ibuprofen
US4996292A (en) * 1989-06-30 1991-02-26 Fox Sidney W Self-sealing artificial skin comprising copoly-alpha-amino acid
JP2911496B2 (en) 1989-09-11 1999-06-23 帝國製薬株式会社 Highly absorbable vaginal agent containing bioactive polypeptide
US5271961A (en) 1989-11-06 1993-12-21 Alkermes Controlled Therapeutics, Inc. Method for producing protein microspheres
US5216124A (en) 1989-12-15 1993-06-01 G. D. Searle & Co. Substituted cyclic tetrapeptides
US5126147A (en) 1990-02-08 1992-06-30 Biosearch, Inc. Sustained release dosage form
FR2658076B1 (en) 1990-02-12 1992-06-12 Sanofi Sa COSMETIC COMPOSITION CONTAINING COPOLYMERS OF AMINO ACIDS, USEFUL AS A MOISTURIZING AGENT.
JP3249147B2 (en) 1990-06-01 2002-01-21 キリン−アムジエン・インコーポレーテツド Oral preparation containing bioactive protein
CA2046830C (en) 1990-07-19 1999-12-14 Patrick P. Deluca Drug delivery system involving inter-action between protein or polypeptide and hydrophobic biodegradable polymer
US5451410A (en) 1993-04-22 1995-09-19 Emisphere Technologies, Inc. Modified amino acids for encapsulating active agents
JPH05239021A (en) 1990-09-04 1993-09-17 Microbial Chem Res Found New actinonin derivative
US5418010A (en) 1990-10-05 1995-05-23 Griffith Laboratories Worldwide, Inc. Microencapsulation process
DE4033419A1 (en) 1990-10-20 1992-04-23 Wolman Gmbh Dr POLYMOUS NITROGEN COMPOUNDS AND METAL FIXING SAEURS CONTAINING WOOD PROTECTION AGENTS
US5137892A (en) 1990-12-12 1992-08-11 Abbott Laboratories Quinoline, naphthyridine and pyridobenzoxazine derivatives
US5244653A (en) 1991-05-01 1993-09-14 Isp Chemicals Inc. Glycine anhydride dimethylol as a biocide and preservative
US5250236A (en) 1991-08-05 1993-10-05 Gasco Maria R Method for producing solid lipid microspheres having a narrow size distribution
ZA93929B (en) 1992-02-18 1993-09-10 Akzo Nv A process for the preparation of biologically active materialcontaining polymeric microcapsules.
US5352461A (en) 1992-03-11 1994-10-04 Pharmaceutical Discovery Corporation Self assembling diketopiperazine drug delivery system
US5310535A (en) 1992-04-24 1994-05-10 The Dow Chemical Company Carboxamide modified polyamine chelators and radioactive complexes thereof for conjugation to antibodies
US5439686A (en) 1993-02-22 1995-08-08 Vivorx Pharmaceuticals, Inc. Methods for in vivo delivery of substantially water insoluble pharmacologically active agents and compositions useful therefor
US5362478A (en) 1993-03-26 1994-11-08 Vivorx Pharmaceuticals, Inc. Magnetic resonance imaging with fluorocarbons encapsulated in a cross-linked polymeric shell
EP0696208B1 (en) * 1993-04-22 2001-08-22 Emisphere Technologies, Inc. Oral drug delivery compositions
ATE171715T1 (en) 1993-04-23 1998-10-15 Rhodia Chimie Sa POLYANHYDROASPARAGIC ACID AND ITS BIODEGRADABLE HYDROLYSIS PRODUCTS

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EP0696208A1 (en) 1996-02-14
IL109403A0 (en) 1994-07-31
DE69428040T2 (en) 2002-05-29
JP4361601B2 (en) 2009-11-11
EP1025840B1 (en) 2005-06-29
ES2163444T3 (en) 2002-02-01
ES2244367T3 (en) 2005-12-16
DE69434418D1 (en) 2005-08-04
AU6819294A (en) 1994-11-08
WO1994023767A1 (en) 1994-10-27
EP1077070A3 (en) 2001-05-23
CA2160693A1 (en) 1994-10-27
ATE204467T1 (en) 2001-09-15
EP1077070A2 (en) 2001-02-21
EP1025840A2 (en) 2000-08-09
DE69434418T2 (en) 2005-12-22
US5766633A (en) 1998-06-16
JP2006273875A (en) 2006-10-12
JP4657164B2 (en) 2011-03-23
EP0696208B1 (en) 2001-08-22
EP1025840A3 (en) 2000-08-30
JPH08509474A (en) 1996-10-08
EP0696208A4 (en) 1997-01-29
ATE298561T1 (en) 2005-07-15
DE69428040D1 (en) 2001-09-27

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