CA2094974C - Stable porfimer sodium compositions and methods for their manufacture - Google Patents
Stable porfimer sodium compositions and methods for their manufactureInfo
- Publication number
- CA2094974C CA2094974C CA002094974A CA2094974A CA2094974C CA 2094974 C CA2094974 C CA 2094974C CA 002094974 A CA002094974 A CA 002094974A CA 2094974 A CA2094974 A CA 2094974A CA 2094974 C CA2094974 C CA 2094974C
- Authority
- CA
- Canada
- Prior art keywords
- composition
- porphyrin
- hematoporphyrin
- sodium hydroxide
- ester
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Lifetime
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Classifications
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/40—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with one nitrogen as the only ring hetero atom, e.g. sulpiride, succinimide, tolmetin, buflomedil
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K41/00—Medicinal preparations obtained by treating materials with wave energy or particle radiation ; Therapies using these preparations
- A61K41/0057—Photodynamic therapy with a photosensitizer, i.e. agent able to produce reactive oxygen species upon exposure to light or radiation, e.g. UV or visible light; photocleavage of nucleic acids with an agent
- A61K41/0071—PDT with porphyrins having exactly 20 ring atoms, i.e. based on the non-expanded tetrapyrrolic ring system, e.g. bacteriochlorin, chlorin-e6, or phthalocyanines
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
Abstract
A storage stable porfimer sodium (polyhematoporphyrin ether/ester) composition useful in the photodynamic therapy of cancer and other conditions is provided. The composition is characterized in that the percentage of ester linked porphyrin oligomers is less than 10% of the composition. Improved processes for preparing such compositions are also provided wherein acetylated hematoporphyrin is treated with alkali for sufficient time and temperature to reduce the amount of porphyrin oligomers joined by ester linkages to less than 10% of the composition.
Description
--2094~74 Title: STABLE PORFIMER SODIUM COMP08ITION8 ~ND METHOD8 FOR TH~IR MANUFACT~R~
BACRGRO~ND OF THE l~.v~,lON
1. Field of the Invention The invention relates to ~torage stable porfimer sodium compositions u~eful in photodynamic therapy and improved method~ for their manufacture.
BACRGRO~ND OF THE l~.v~,lON
1. Field of the Invention The invention relates to ~torage stable porfimer sodium compositions u~eful in photodynamic therapy and improved method~ for their manufacture.
2. DescriPtion of the Related Art The u~e of porphyrin compound~, and in particular hematoporphyrin and its derivative mixture hematoporphyrin derivative (HPD), have been known for some time to be useful syQtemically when combined with irradiation, for the treatment and diagnosis of malignant cells. The porphyrins appear to "naturally"
localize in malignant tissue where they absorb light at certain wavelengths when irradiated, providing a mean~
to detect the tumor by the location of the fluore~-cence. Accordingly, preparations cont~; n~ ng the porphyrins are useful in the diagnoQis an~ detection of such tumor tissues. tSee e.g. "Porphyrin Photo~ensiti-zation~', Ressel, D., et al., ed~. (1983) Plenum Press).
In addition, the porphyrin~ al~o have the capability of exhibiting a cytotoxic effect on the cells or other tis~ue~ in which they are localized when irradiated at the appropriate wavelength. (See, e.g., Diamond, I., et al., Lancet (1972) 2: 1175-1177; Dougherty, T.J. et al., ~The Science of Photo Medicine~ (1982)~ J.D. Regan r & J.A. Parri~h, eds., pp. 625-638; Dougherty, ~.J., et al., "Cancer: Prineiple~ and Practice of Oneology"
(1982) V.T. DeVita Jr., et al., edq., pp. 1836-1844).
It has been postulated that the cytotoxie effeet of the S porphyrin~ is due to the formation of ~inglet oxygen upon irradiation (Wei~haupt, R.R., et al., Caneer Research, (1976) 36: 2326-2329). The suceesQful treatment of AIDS-related oral Rapo~ 8areoma with a purified form of HPD, Photofrin~ porfimer sodium, was deseribed in ~ehweitzer, V.G. et al., OtolaryngologY --Head and Neek 8urgery, ~1990) 102: 639-649.
In addition to ~ystemie use for the treatment and diagnoqis of tumors, the porphyrins ean be used in a variety of other therapeutie applieations. For example, photosensitizer~ are useful in the deteetion and treatment of artheroselerotic plaqueq a~ disclosed in U.S. Patent No. 4,517,762 and 4,577,636. U.8.
Patent Nos. 4,500,507 and 4,485,806 deQcribe the use of radiolabeled porphyrin eompounds for tumor imaging.
Porphyrin compounds have also been used topieally to treat various skin di~eases as diselosed in ~.S. Patent No. 4,753,958.
A number of porphyrin photo~ensitizer preparations have been disclosed for therapeutie applieations. A photoQensitizer preparation widely used in the early ~tage-q of photodynamie therapy both for detection and tre~tment was a crude derivative of hematoporphyrin, also called hematoporphyrin derivative (HPD) or Lipson derivative, prepared a~ de~eribed by Lipson et al., J. Natl. eancer In~t. (1961) 26: 1-8. A
purified form of the active component(s) of HPD was prepared by Dougherty and co-workers by adju~tment of the pH to cause aggregation and reeovery of the aggregate, as diselosed in U.S. Patent~ ~,649,151, 4,866,168, 4,889,129 and 4,932,934. A purified form of this product haQ been used clinically under the 2Q~4974 trademark Photofrin~ porfimer sodium ~al~o referred to as polyhematoporphyrin ether/e~ter~).
Porfimer sodium i~ prepared by following the method of Lip~on a~ modified by Dougherty et al. In brief, hematoporphyrin is fir~t treated with a mixture of sulfuric and acetic acid~ which produces a mixture of mono- and di-acetate of hematoporphyrin (Hpl and it~
dehydration products, hydroxyethylvinyldeutero porphyrin (Hvd) and protoporphyrin. ThiQ mixture upon treatment with 0.1 N NaOH undergoe~ hydroly~i~ and coupling to produce hematoporphyrin derivative (~pd) which is a mixture of Hp, Hvd, protoporphyrin and a higher molecular weight fraction consi~ting of a mixture of porphyrin oligomers. In the Dougherty et al. disclo~ure, this higher molecular weight fraction was separated by adjustment of the pH to around 9.5 and removal of the resulting aggregate by membrane filtration. See U.8. Patent No. 4,649,151.
The structure of porfimer sodium and the nature of linkage~ joining the porphyrin unit~ has been the subject of discu~sion for guite some time. Bonnett and Berenbaum, Adv. Exp. Biol. Med. 160, 2~1 (1983) proposed that the ~tructure of the tumor localizing component might be a dimer or oligomer linked by ether or carbon to carbon covalent bond~. Dougherty et al., Prog. Clin. Biol. Re~., (1984) 170: 301-31~, presented evidence from fast atom bomb rdment ma~s ~pectro~cGpy (FAB-MS) and NMR spectroscopy that the localizing component~ were most likely the i~omer~ of dihematopor-phyrin ether. Recently Ressel and coworker~, CancerRes. 47, 4642 (1987) have ~uggested that the tumor localizing component~ of porfimer ~odium might be porphyrin dimer~ with e~ter and ether linkage~.
However, the presence of a polymeric mixture wa~ not ruIed out. Recently it has been reported th~t hematoporphyrin dimer, trimer and their dehydration product~ with ether linkage~ are biologi¢ally active and are likely active components in porfimer sodium, Pandey et al., Tetrahedron Lett. 29: 4567 ~1988).
Hematoporphyrin dimer with e~ter linkago wa~ found to be biologically inactive and i~ only a minor component of porfimer ~odium, Pandey et al., Cancer Re~. 49: (8) 2042-2047 (1989). It i~ now believed, from fa~t atom bombardment ma~ ~pectral analy~i~ of porfimer ~odium, that it i~ in fact a complex mixture of oligomer~
having from two up to eight porphyrin unit~ linked together and that the porphyrin unit~ are hematopor-phyrin, hydroxyethylvinyl-deuteroporphyrin and protoporphyrin unit~. The porphyrin unit~ are linked with ether and/or e~ter bond~ a~ generally exemplified -in ~tructure I, below:
II-O~C(CN~ c~ e~ c~ e~t)lco~
~ ~ ~_ ~ .",eo,~
--~<~ N ~ e ~1/ ~ C N
C~ ~e~e~ ell~ eN~ t _ -- n In the foregoing ~tructure~, n i~ an integer from O to 6 and R i~ hydroxyetbyl or vinyl. It i~ understood that the compo~ition includes a combination of oligomer~ of varying chain length, that i~, a combina-tion of compound~ where n i~ 0, 1, 2, 3, ~, 5 or 6 (dimer~, trimer~, tetramer~, etc.). In addition, the compo~ition include~ a combination of hematoporphyrin, hydroxyethylvinyl-deuteroporphyrin, and protoporphyrin units (i.e. R i~ hydroxyethyl and/or vinyl).
The porphyrin unit~ may be linked together in different ring orientation~, depending on whether ether or ester bond~ are involved. Referring to the structure below:
N~o2C(CH2)2 CH3 N~02C(CH2)2 ~\ /~ CH3 ~_ R CH~
where the ring~ of th~ porphyrin are labelled A, B, C
and D, the ether lin~ed porphyr~n~ may be ioined only with A-A, A-B, B-A, or B-B orientation~. B~ter lin~ed porphyrin~ may be joined with the A-C, A-D, B-C or B-D
orientation~.
Heretofore, the u~e of porfimer ~odium a~ a therapeutic or diagno~tic agent ha~ been hindered by the relative instability of the product. Porfimer sodium prepared by the prior art proce~ degrade~
rapidly when exposed to heat and i~ ~table for only a few hours at room temperature. Accordingly, the solution is generally kept frozen and thawed immediately prior to use.
The use of such a frozen solution ha~ many obvious ai~advantages. Because it has to be ~ept frozen, it must be shipped and stored in a frozen state, necessitating the use of ~pecial freezing conditions. For example, the product mu~t be shipped in special containers using dry ice or the li~e as a refrigerant. This is a major drawback, adding to the cost and logistics of using the product. At the point of use, the frozen solution must be stored at -20C, which is below the operating temperatures of some freezers thereby necessitating ~pecial freezer eguipment. In addition, the frozen product must undergo a thawing period and is therefore not useable immediately with a patient.
Various methods have been proposed to overcome that instability of the product. ~.8. Patent 5,059,619 discloses a ~table, freeze-dried preparation of porfimer sodium which is obtained by freeze-drying an aqueous solution of the photosensitizer compo~ition.
This preparation is reconstituted prior to administra-tion. ~ai et al., U.g. Patent ~,882,234, di~closes a storage stable composition which is obtained by heat treating a buffered pH 5-9 aqueous ~olution of hematoporphyrin at a temperature of 60C to about 120C
for 5-100 hours. The resulting product appears to have a different composition than porfimer sodium and is stated to contain at lea~t 80% by weight of itQ
porphyrin~ as ester lin~ed oligomers where the oligomeric chain is an average of about 3 to 7.
It is an object of the present invention to provide porfimer sodium compositions which exhibit enhanced thermal and temporal stability as compared to - preparations shown in the art, and which maintain high 2094~74 levels of activity as photodynamic therapeuti¢ and diagnostic agents. It i~ a further ob~ect of this invention to provide processes for producing such materials and methods of using such materials.
s 8~MMARY OF TRE INV~NTION
It has been found that a composition of porfimer sodium which exhibits enhanced stability may be obtained by ta~ing the product formed from the reaction of hematoporphyrin with acetic/sulfuric acid, (hereinafter referred to as "the acetylated hematoporphyrin composition~'), and reacting it with al~ali at such conditions to reduce the amount of porphyrin oligomers joined by ester lin~ages to less than 10%, preferably 4-5% tas shown by high performance liquid chromatography (HPLC) in con~unction with base hydrolysis). The resulting product has enhanced thermal and temporal stability and therefore a greater shelf-life.
The porfimer sodium compositions of the present invention may be defined as a biologically active composition being flourescent, photosensitizing and having the capability of localizing in ~nd being retained in tumor tissue as compared to normal tissues which composition comprises a mixture of porphyrin oligomers, said oligomers comprised of two to eight porphyrin molecules covalently lin~ed by ether and ester bonds wherein at least one porphyrin molecule has the formula:
lio~e~CR~
~o~
wherein R is hyroxyethyl or vinyl and wherein the bond shown forms said covalent linkage; and the pharmaceutically acceptable salts thereof; with the proviso that the percentage of ester linkages in said composition are less than 10% of the linkages in the composition as shown by HPLC in conjunction with base hydrolysis.
The resulting porfimer sodium compositions of the present invention are characterized, as stated, in that they contain less than 10% of their oligomeric linkages as ester bonds as shown by base hydrolysis on HPLC assay. These compositions are prepared by employing processes which allow the ester oligomeric linkages to hydrolyze in the reaction process of the acetylated hematoporphyrin composition with alkali, leading to a higher percentage of ether linkages in the product. The ester content of porfimer sodium can be characterized by utilizing HPLC in conjunction with exhaustive base hydrolysis. In this procedure, the product is assayed before and after treatment with base which hydrolyzes the ester bonds. As stated, the product exhibits enhanced stability as compared to the composition of the prior art.
The present invention can therefore be described by the processes by which the porfimer sodium oligomer products are formed. Such B
a process includes the step of reacting the acetylated hematoporphyrin composition with alkali for sufficient time and B
temperature to hydrolyze the e~ter lin~age~ of the oligomeric product~ formed such that there are le~
than 10% by weight ester-lin~ed oligomer~ in the composition. Alternatively, the acetylated hematopor-phyrin composition can be reacted with alk~li,preferably ~odium hydroxide, and the re~ulting product i~ held at an al~aline pH for sufficient timo to hydrolyze the ester lin~age~.
In a preferred embodiment, the prep~ration lo proce~ comprise~ the re~ction of the acetylated hematoporphyrin composition with O.lN-0.2N ~odium hydroxide at room temperature for 16-2~ hour~. The pH
of the ~olution i~ adjusted to 9.~-9.6 with hydrochloric acid and purified by diafiltration with a membrane filter to eliminate low molecular weight components.
In additional aspects, this in~ention provides compositions cont~;ning the predominantly ether lin~ed oligomer composition~ and their use in photodynamic therapy and diagnosis.
--lO- 2094974 DETAI~ED DE8CRIPTION
BRIEF DE8CRIPTION OF TB DRA~ING~
In the specification and claims, reference will be made to the accompanying drawings in which:
FIG. 1 is a graph showing the stability of the compositions of the present invention over time at 2-4C as compared to the composition of the art.
FIG. 2 is a graph showing the stability, over time at 4C, of compositions prepared by reacting the acetylated hematoporphyrin composition with al~ali for a period of 24 hours as compared with a composition of the art.
DE8CRIPTION OF T B PREFERRED EMBODI~NT8 The ~tarting material employed in tho present invention is hematoporphyrin dihy~rochloride.
Hematoporphyrin dihydrochloride is derived from tho naturally occurring compound, heme, and i9 available commercially from for example, Aldrich Chemical Company.
The material i8 availablo in its protonated ~Cl salt form as a dry powder.
2s In accordance with tho pres~nt invention, the hematoporphyrin dihydrochloride starting matorial is reacted with acetic acid/sulfuric acid a~ taught by Lipson et al., J. Natl. Cancer Inst. 26: 1-8 (1961) and Dougherty et al., U.~. Patent 4,649,151 to produce the acetylated hematoporphyrin composition. This compo~ition is then reacted with al~ali, preferably 0.lN-0.2N sodium hydroxide, for ~ufficient ti~o and at sufficient temperature to hydrolyze tho ester lin~ages ~uch that there is less than 10%, proferably lo~s than 4-5% esters in the composition.
In one embodiment, the acetylated hematopor-phyrin i~ reacted with O.lN ~odium hydroxide for at le~st ~n hour at room temperature. The solution is purified using room temperature diafiltration and S stored at 4C from 14 to 21 day~. The m~terial is then repurified u~ing a diafiltration procedure.
In an alternate procedure, the acetylated hematoporphyrin is dissolved in either O.lN or 0.2N
~odium hydroxide and allowed to react for lC to 24 hours at room temperature. The solution is then purified using a room temperature diafiltration procedure or using a diafiltration procedure at 35-40 C.
U~ing an additional method, the acetylated hematoporphyrin is dissolved in either O.lN or 0.2N
sodium hydroxide and allowed to react for one hour at 35-45C. The solution is then purified at room temperature or at 35-45C using a diafiltration procedure.
Alternatively, the acetylated hematoporphyrin i~ di~olved in O.lN sodium hydroxide and allowed to react for one hour at room temperature. Additional sodium hydroxide is then added to a concentration of 0.2N and held there for 10-24 hours or a lN concentra-tion and held for 2 hours. The ~olution is then purified at room temperature using the diafiltration procedure.
In an alternative procedure, acetylated hematoporphyrin i~ di~solved in O.lN sodium hydroxide and allowed to react for one hour at room temperature.
The pH of the ~olution is adjusted to 9.6 with hydrochloric acid and the resulting mixture held for 3 days at room temperature.
The improvement~ over the Dougherty proces~
in each of the aforementioned variation~ re~ult~ in final solutions that are more stable, thu~ allowing -12- 20~4974 ~torag- in the liguid ~tate at l-~C over an extended p-riod of time At pr-sent, th- pr-ferr-d embodim-nt for preparing th- ~table porfimer ~odium compo~ition i~ to react th- acetylat-d hematoporphyrin with O lN ~odium hydroxide and allowing th- reaction to occur for 16-2~
hours The ~olution i~ th-n purified by diafiltration at room temperature The ~olution of the ac-tylated hematopor-phyrin compo~ition is an agu-ou~ ~olution The aqueous alkali ~olution i~ generally in a concentration of O lN-0 2N, but concentrations of up to lN may be used Generally, the ratio of acetylated hematoporphyrin to alkali ~olution i8 1: 50 The time and temperature for the reaction are related Higher temperature~ employ ~horter time~
while lower temperature~ reguir- longer period~ of time to hydrolyze the ester~ Th- concentration of alkali i~ al~o related, in that ~tronger ~lkali concentration~
require ~horter r-action times The temperature is generally moderate, fro~ room temperature to temp-rature~ of 40-~5C ~igher temperature~ are not preferred since degradation of the productQ tends to occur at higher temperature~ The time, temp-rature and ~trength of the alkali Qhould be adequate to hydrolyze the ester bond~ such that about 90% or greater of the oligomer~ are ether linked oligomer~
Thi~ conversion can be monitored by ~uch technique~ as exhaustive ba~e hydrolyQis followed by HP~C ~high performance liguid chromatography) analy~i~ Examples of time~ and temperature~ which give the de~ired re~ults are about 4C for 1~-21 day~, roo~ t-~perature for 16-2~ hour~, 35-~5C for on- hour, room t-~perature for one hour, adju~tment of the pH to 9 6 and holding for 3 days, then purification at room temperature -The oligomer product of the present invention can be characterized in terms of its chemicA~l ma~e-up and properties. Although not understood with certainty, it is believed that the produ~At is a mixture of oligomers in which up to eight porphyrin units which are hematoporphyrin, hydroxyethylvinyldeuteroporphyrin and protoporphyrin units are covalently linked together by ether or ester bonds as described on page ~-5 of this application. It is a property of the present invention that no more than 10% of the composition consists of ester linked oligomers.
The product may be characterized by determining the ester content of the composition using HPLC in conjunction with exhaustive ba~e hydrolysis.
In this procedure, the product is assayed on HPLC to establish a baseline, then subjected to exhaustive treatment with base, such as lN sodium hydroxide for 12-18 hours to hydrolyze all of the ester bonds. The resulting product is then assayed once more on XPLC.
By comparing the HPLC chromatogram before and after base hydrolysis one can determine the percentage ester content in the original composition. As stated, it is a property of the claimed composition that it contains no more than 10% ester lin~ed oligomers when analyzed using this procedure.
Without intending to be bound to any particular theory of how the present invention operates, it is now believed by the present inventors that the invention relies on a series of reactions to yield the composition of the invention. In the fir~t reaction, the acetylated hematoporphyrin composition forms oligomeric lin~ages when reacted with al~ali.
Some of the oligomers formed are ether lin~ages and some are esters. If the reaction i~ allowed to continue for appropriate time and under appropriate conditions, the ester lin~ages hydrolyze and the resulting eompositionQ formed is then predominantly ether linked oligomers whieh show enhaneed stability.
A~ stated above, the compo~itions of the pre~ent invention show enhaneed stability. ~able 1 ~how~ a eompari~on of the purity of the produets produeed by the prior art proee~ aiselosed by U.8.
Patent 4,649,151 (~the Dougherty proees~) ver~u~ three of the variation~ whieh inelude (a) keeping the reaetion of the aeetylated hematoporphyrin and O.lN
sodium hydroxide at room temperature for 24 hour followed by diafiltration at room temperature: (b) ~eeping the reaetion of the aeetylated hematoporphyrin and O.lN ~odium hydroxide for 1 hour at ~0-45C; or te) keeping the reaetion of the aeetylated hematoporphyrin -and O.lN sodium hydroxide at room temperature for 1hour, adjusting the pH to 9.6 with lN hydroehloric aeid, holding for 3 days followed by diafiltration at room temperature. AS is evident from looking at the data in Table 1, analysi~ of the resulting produet in eaeh ease ~how~ that over time while being stored at 2-4C the produet of the Dougherty Proee~s degrades from 92% to 86% while the products obtained with each of the various improvements results in produet~ with only minor degradation.
2s Table I: ~ Porfimer Sodium vs. Storage Time at 2C-4C
Storage Time Dougherty 24-Hour High pH 9.6 (Days) Process Hold Temperature 3 Days 0 92.1 90.8 92.9 91.0 1 9~.3 2 90.3 4 89.8 91.0 6 91.6 7 88.8 93.3 8 91.3 9 87.9 11 87.5 14 87.2 90.6 92.8 92.6 18 86.5 89.4 21 92.0 92.0 23 86.0 28 85.8 91.3 92.3 92.0 34 90.2 92.4 49 90.1 Dougherty Process: 0.lN NaOH 1 hour, purification at RT
24-Hour Hold: 0.lN NaOH 24 hour~, purification at RT
High Temperature: 0.1N NaOH 1 hour at 40-45C, purification at 40-45C
pH 9.6, 3-Day Hold: 0.1N NaOH 1 hour, pH to 9.6 w/lN HCl, 3-day hold, purification at RT
~
A~ further ~hown in Figure 1, the product of the Dougherty Proees~ h~ a con~tant lo~ of purity over time while each of the product~ obtained through the enhaneed proceQ~e~ (a), ~b) and (e) above ~how~
enhanced -Qtability at 2-~C.
AQ ~hown graphieally in Figure 2 the product obtained in eaeh of several preparation~ u~ing the preferred proeedure of reaeting the aeetylated hematoporphyrin and O.lN ~odium hydroxiae at room temperature for 24 hour-Q followed by diafiltration at room temperature retainQ produet ~tability over time ver~u~ produet prepared by the Dougherty proee~ Q .
The porfimer ~odium oligomer produet of thi~
invention ean be formulated into pharmaeeutieal preparations for injeetion. Sueh preparationQ can eontain from 0.1 to about 3.0 gramQ per liter of the porfimer -Qodium material in a pharmaeeutieally aeceptable nonpyrogenie aqueou~ injection vehicle.
Typical preparation~ have pH'~ between about 7.0 and about 8.1 and are iQotonic. They al~o are ~ub~tan-tially free of any solid contaminantQ. The eoncentra-tion, pH and ionic ~trength of the preparation~ can be adjusted by removing water by ultrafiltration, by adding liquid~ ~ueh a~ ~terile water for injeetion, ~terile ~aline for injeetion or baeterio~tatic water for injeetion a~ the~e liquid~ are defined in the book Remington~ Pharmaeeutieal Seienee~, 1985, Maek Publishing Company, 1985, by adding pH adju~ting material~ ~ueh a~ U.S.P. sodium hydroxidQ or ~.8.P.
phosphorie aeid, or the like. The pharmaeeutieal preparations are then paekaged under ~terile eondition~
in ~uitable vialQ, bottle~, prefilled ~yringe~, ana the like for admini~tration via intravenou~ infu~ion or intramu~eular injeetion to patient~.
The porfimer Qodium oligomer produet~ of the invention may be u~ed in photodynamic therapy or d~agno~is in the manner which i~ di~clo~ed for such porphyrin based product~. The amount of porfimer ~odium oligomer product administered to the patent can range from about 0.1 mg to 10 mg per ~ilogram of body weight with do~age~ of 1 to 2 mg/~g being preferred.
The invention will now be further de~cribed with reference to the following Example~. The~e are presented ~olely to ma~e clear preferred embodiments of the pre~ent invention and are not to be con~trued a~
limiting the ~cope of the invention which i~ as defined by the appended claim~.
~xample 1 Acetylated hematoporphyrin (1 part by weight) i5 dissolved in O.lN ~odium hydroxide (50 part~ by volume) and stirred for one hour at room temperature.
After the ~tir period the ~olution i~ adju~ted to pH
9.4 to 9.6 with lN hydrochloric acid. It i8 filtered through a 5~m filter and then concentrated to 1/4(12.5 part~) of its original volume in an ultrafilter with 10,000 molecular weight cut off membrane~. The solution i~ then purified via diafiltration maint~;~ing con~tant volume with 120 volume~ of water and ~eeping the pH at 9.~ to 9.6 with O.lN ~odium hydroxide. Thi~
i~ also done at room temperature. After the purification, the ~olution i~ removed from the ultrafilter, diluted to 3/8(18.8 parts) of it~ original volume and pH adju~ted to 7.5 to 7.7 with lN
hydrochloric acid. The ~olution i~ then ~tored at 4C
for 14 to 21 day~. After ~torage, the ~olution i~ pH
adju~ted to 9.4 to 9.6 with O.lN ~odium hydroxide and concentrated to 1/4(12.5 part~) of it~ original volume.
The solution i8 then repurified a~ above. The ~olution iq diluted to 3/8(18.8 part~) of it~ original volume and pH adju~ted to 7.5 to 7.7 with lN hydrochloric acid. The solution i~ then analyzed and, if nece~ary, it i~ adjusted to between 13 to 18 mg/mL by the addition of water The solution i~ then filtered through a 0 22~m filter into bottles for storage at 1C
to 4C to await further processing ~xample 2 Acetylated h-matoporphyrin ~1 part by weight) is dissolved in O lN or 0 2N sodium hydroxid- ~50 parts by volume) and stirr-d for 16 to 24 hours at room te~perature After th- stir period th- solution is adjusted to p~ 9 4 to 9 6 with lN hydrochloric acid It is filtered through a 5~ filter and th-n concentrated to 1/4~12 5 part~) of its original volume in an ultrafilter with 10,000 molecuiar w-ight cut off membranes The solution i~ then purifi-d via diafiltration at room temperature or at 35C to 45C, maintaining constant volume with 120 volumes of water and keeping the p~ at 9 4 to 9 6 with O lN sodium hydroxide After tho purification, th- solution is removed from the ultrafiltor, diluted to 3/8(18 8 part~) of it~ original ~olu~e and pH adjusted to 7 5 to 7 7 with lN hydrochloric acid The solution is then analyzed and, if necessary, it is ad~ust-d to between 13 to 18 mg/m~ by the addition of ~ater Th- ~olution is then filt-red through a 0 22~m filter into bottles for storage at 1C to 4C to a~ait further processing ~xamDl- 3 Acetylated hematoporphyrin (1 part by weight) is dissolved in O lN or 0 2N ~odiu~ hydroxid- (50 parts by volum-) and stirr-d for one hour at 3S to 45C
After the stir period the solution i~ ad~ust-d to pH
9 4 to 9 6 with lN hydrochloric aci~ It i~ filter-d through a 5~m filt-r and then conc-ntratsd to 1/~(12 5 parts) of its original volume in an ultrafilter with 10,000 molecular weight cut off me~bran-s Th-solution is then purified via diafiltration at room temperature or at 35 to 45C, maint-in~ng constant volume with 120 volum-s of water and ~eeping the p~ at 9.4 to 9.6 with o.lN sodium hydroxide. After the purification, the _olution is removed from the ultrafilter, diluted to 3/8(18.8 part_) of itQ original volum- and pH adjusted to 7.5 to 7.7 with lN
s hydrochlori¢ acid. The solution is then analyzed and, if nece_sary, it is adjuQted to between 13 to 18 mg/mL
by the addition of water. The solution is then filtered through a 0.22~m filter into bottleQ for storage at 1C to 4C to await further processing.
~xample 4 Acetylated hematoporphyrin ~1 P~rt by weight) i~ di~Qolved in 0.lN sodium hydro~ide ~50 parts by volumo) and stirred for 1 hour at room temperature.
Sodium hydroxide is then added to give an overall concentration ~based on initial NaO~ plus this addition) of either l.lN ~2 parts NaOH) or 0.2N ~0.2 parts NaOH). ThiQ solution is then stirred for 2 hours (l.lN NaOH) or 24 hours ~0.2N NaOH). After the stir period the solution is ad~usted to pH 9.4 to 9.6 with lN hydrochloric acid. It is th-n filtered through a 5~m filter and then conc-ntrated to 1/4~12.5 parts) of it~ original volume in an ultrafilter with 10,000 molecular weight cut off membranes. The solution i_ then purified via diafiltration maintaining constant volume with 120 volume~ of water and keeping the p~ at 9.4 to 9.6 with 0.lN sodium hydro~id-. This is also done at roo~ temperature. Aft-r the purification, the solution is removed from th- ultrafilt-r, diluted to 3/8~18.8 parts) of its original volume and p~ adjusted to 7.5 to 7.7 with lN hydrochloric acid. The solution is then analyzed and, if n-c-ssary, it is ad~usted to between 13 to 18 mg/~L by tho addition of water. The solution is th-n filtered through a 0.22~m filter into bottles for storage at ~C to 4C to await further proce~sing.
~xample 5 Acetylated hematoporphyrin (1 part by weight) is dissolved in O.lN sodium hydroxide (50 part~ by volume) and stirred for one hour at room temperature.
After the stir period the ~olution i8 adju~ted to pH
9.4 to 9.6 with lN hydrochloric acid and i~ held at room temperature for 3 day~. After the holding period, the solution i~ filtered through a S~m filter and then concentrated to 1/4(12.5 part~) of it~ original volume in an ultrafilter with 10,000 molecular weight cut off membrane~. The solution i~ then purified via diafiltration maintaining con~tant volume with 120 volume~ of water and ~eeping the pH at 9.4 to 9.6 with o.lN sodium hydroxide. Thi~ i~ al~o done at room temperature. After the purification, the solution i~
removed from the ultrafilter, diluted to 3/8~18.8 parts) of it~ original volume and pH adju~ted to 7.5 to 7.7 with lN hydrochloric acid. The ~olution i~ then analyzed and, if necessary, it is adju~ted to between 13 to 18 mg/mL by the addition of water. The ~olution is then filtered through a 0.22~m filter into bottle~
for storage at 1C to 4C to await further proce~ing.
localize in malignant tissue where they absorb light at certain wavelengths when irradiated, providing a mean~
to detect the tumor by the location of the fluore~-cence. Accordingly, preparations cont~; n~ ng the porphyrins are useful in the diagnoQis an~ detection of such tumor tissues. tSee e.g. "Porphyrin Photo~ensiti-zation~', Ressel, D., et al., ed~. (1983) Plenum Press).
In addition, the porphyrin~ al~o have the capability of exhibiting a cytotoxic effect on the cells or other tis~ue~ in which they are localized when irradiated at the appropriate wavelength. (See, e.g., Diamond, I., et al., Lancet (1972) 2: 1175-1177; Dougherty, T.J. et al., ~The Science of Photo Medicine~ (1982)~ J.D. Regan r & J.A. Parri~h, eds., pp. 625-638; Dougherty, ~.J., et al., "Cancer: Prineiple~ and Practice of Oneology"
(1982) V.T. DeVita Jr., et al., edq., pp. 1836-1844).
It has been postulated that the cytotoxie effeet of the S porphyrin~ is due to the formation of ~inglet oxygen upon irradiation (Wei~haupt, R.R., et al., Caneer Research, (1976) 36: 2326-2329). The suceesQful treatment of AIDS-related oral Rapo~ 8areoma with a purified form of HPD, Photofrin~ porfimer sodium, was deseribed in ~ehweitzer, V.G. et al., OtolaryngologY --Head and Neek 8urgery, ~1990) 102: 639-649.
In addition to ~ystemie use for the treatment and diagnoqis of tumors, the porphyrins ean be used in a variety of other therapeutie applieations. For example, photosensitizer~ are useful in the deteetion and treatment of artheroselerotic plaqueq a~ disclosed in U.S. Patent No. 4,517,762 and 4,577,636. U.8.
Patent Nos. 4,500,507 and 4,485,806 deQcribe the use of radiolabeled porphyrin eompounds for tumor imaging.
Porphyrin compounds have also been used topieally to treat various skin di~eases as diselosed in ~.S. Patent No. 4,753,958.
A number of porphyrin photo~ensitizer preparations have been disclosed for therapeutie applieations. A photoQensitizer preparation widely used in the early ~tage-q of photodynamie therapy both for detection and tre~tment was a crude derivative of hematoporphyrin, also called hematoporphyrin derivative (HPD) or Lipson derivative, prepared a~ de~eribed by Lipson et al., J. Natl. eancer In~t. (1961) 26: 1-8. A
purified form of the active component(s) of HPD was prepared by Dougherty and co-workers by adju~tment of the pH to cause aggregation and reeovery of the aggregate, as diselosed in U.S. Patent~ ~,649,151, 4,866,168, 4,889,129 and 4,932,934. A purified form of this product haQ been used clinically under the 2Q~4974 trademark Photofrin~ porfimer sodium ~al~o referred to as polyhematoporphyrin ether/e~ter~).
Porfimer sodium i~ prepared by following the method of Lip~on a~ modified by Dougherty et al. In brief, hematoporphyrin is fir~t treated with a mixture of sulfuric and acetic acid~ which produces a mixture of mono- and di-acetate of hematoporphyrin (Hpl and it~
dehydration products, hydroxyethylvinyldeutero porphyrin (Hvd) and protoporphyrin. ThiQ mixture upon treatment with 0.1 N NaOH undergoe~ hydroly~i~ and coupling to produce hematoporphyrin derivative (~pd) which is a mixture of Hp, Hvd, protoporphyrin and a higher molecular weight fraction consi~ting of a mixture of porphyrin oligomers. In the Dougherty et al. disclo~ure, this higher molecular weight fraction was separated by adjustment of the pH to around 9.5 and removal of the resulting aggregate by membrane filtration. See U.8. Patent No. 4,649,151.
The structure of porfimer sodium and the nature of linkage~ joining the porphyrin unit~ has been the subject of discu~sion for guite some time. Bonnett and Berenbaum, Adv. Exp. Biol. Med. 160, 2~1 (1983) proposed that the ~tructure of the tumor localizing component might be a dimer or oligomer linked by ether or carbon to carbon covalent bond~. Dougherty et al., Prog. Clin. Biol. Re~., (1984) 170: 301-31~, presented evidence from fast atom bomb rdment ma~s ~pectro~cGpy (FAB-MS) and NMR spectroscopy that the localizing component~ were most likely the i~omer~ of dihematopor-phyrin ether. Recently Ressel and coworker~, CancerRes. 47, 4642 (1987) have ~uggested that the tumor localizing component~ of porfimer ~odium might be porphyrin dimer~ with e~ter and ether linkage~.
However, the presence of a polymeric mixture wa~ not ruIed out. Recently it has been reported th~t hematoporphyrin dimer, trimer and their dehydration product~ with ether linkage~ are biologi¢ally active and are likely active components in porfimer sodium, Pandey et al., Tetrahedron Lett. 29: 4567 ~1988).
Hematoporphyrin dimer with e~ter linkago wa~ found to be biologically inactive and i~ only a minor component of porfimer ~odium, Pandey et al., Cancer Re~. 49: (8) 2042-2047 (1989). It i~ now believed, from fa~t atom bombardment ma~ ~pectral analy~i~ of porfimer ~odium, that it i~ in fact a complex mixture of oligomer~
having from two up to eight porphyrin unit~ linked together and that the porphyrin unit~ are hematopor-phyrin, hydroxyethylvinyl-deuteroporphyrin and protoporphyrin unit~. The porphyrin unit~ are linked with ether and/or e~ter bond~ a~ generally exemplified -in ~tructure I, below:
II-O~C(CN~ c~ e~ c~ e~t)lco~
~ ~ ~_ ~ .",eo,~
--~<~ N ~ e ~1/ ~ C N
C~ ~e~e~ ell~ eN~ t _ -- n In the foregoing ~tructure~, n i~ an integer from O to 6 and R i~ hydroxyetbyl or vinyl. It i~ understood that the compo~ition includes a combination of oligomer~ of varying chain length, that i~, a combina-tion of compound~ where n i~ 0, 1, 2, 3, ~, 5 or 6 (dimer~, trimer~, tetramer~, etc.). In addition, the compo~ition include~ a combination of hematoporphyrin, hydroxyethylvinyl-deuteroporphyrin, and protoporphyrin units (i.e. R i~ hydroxyethyl and/or vinyl).
The porphyrin unit~ may be linked together in different ring orientation~, depending on whether ether or ester bond~ are involved. Referring to the structure below:
N~o2C(CH2)2 CH3 N~02C(CH2)2 ~\ /~ CH3 ~_ R CH~
where the ring~ of th~ porphyrin are labelled A, B, C
and D, the ether lin~ed porphyr~n~ may be ioined only with A-A, A-B, B-A, or B-B orientation~. B~ter lin~ed porphyrin~ may be joined with the A-C, A-D, B-C or B-D
orientation~.
Heretofore, the u~e of porfimer ~odium a~ a therapeutic or diagno~tic agent ha~ been hindered by the relative instability of the product. Porfimer sodium prepared by the prior art proce~ degrade~
rapidly when exposed to heat and i~ ~table for only a few hours at room temperature. Accordingly, the solution is generally kept frozen and thawed immediately prior to use.
The use of such a frozen solution ha~ many obvious ai~advantages. Because it has to be ~ept frozen, it must be shipped and stored in a frozen state, necessitating the use of ~pecial freezing conditions. For example, the product mu~t be shipped in special containers using dry ice or the li~e as a refrigerant. This is a major drawback, adding to the cost and logistics of using the product. At the point of use, the frozen solution must be stored at -20C, which is below the operating temperatures of some freezers thereby necessitating ~pecial freezer eguipment. In addition, the frozen product must undergo a thawing period and is therefore not useable immediately with a patient.
Various methods have been proposed to overcome that instability of the product. ~.8. Patent 5,059,619 discloses a ~table, freeze-dried preparation of porfimer sodium which is obtained by freeze-drying an aqueous solution of the photosensitizer compo~ition.
This preparation is reconstituted prior to administra-tion. ~ai et al., U.g. Patent ~,882,234, di~closes a storage stable composition which is obtained by heat treating a buffered pH 5-9 aqueous ~olution of hematoporphyrin at a temperature of 60C to about 120C
for 5-100 hours. The resulting product appears to have a different composition than porfimer sodium and is stated to contain at lea~t 80% by weight of itQ
porphyrin~ as ester lin~ed oligomers where the oligomeric chain is an average of about 3 to 7.
It is an object of the present invention to provide porfimer sodium compositions which exhibit enhanced thermal and temporal stability as compared to - preparations shown in the art, and which maintain high 2094~74 levels of activity as photodynamic therapeuti¢ and diagnostic agents. It i~ a further ob~ect of this invention to provide processes for producing such materials and methods of using such materials.
s 8~MMARY OF TRE INV~NTION
It has been found that a composition of porfimer sodium which exhibits enhanced stability may be obtained by ta~ing the product formed from the reaction of hematoporphyrin with acetic/sulfuric acid, (hereinafter referred to as "the acetylated hematoporphyrin composition~'), and reacting it with al~ali at such conditions to reduce the amount of porphyrin oligomers joined by ester lin~ages to less than 10%, preferably 4-5% tas shown by high performance liquid chromatography (HPLC) in con~unction with base hydrolysis). The resulting product has enhanced thermal and temporal stability and therefore a greater shelf-life.
The porfimer sodium compositions of the present invention may be defined as a biologically active composition being flourescent, photosensitizing and having the capability of localizing in ~nd being retained in tumor tissue as compared to normal tissues which composition comprises a mixture of porphyrin oligomers, said oligomers comprised of two to eight porphyrin molecules covalently lin~ed by ether and ester bonds wherein at least one porphyrin molecule has the formula:
lio~e~CR~
~o~
wherein R is hyroxyethyl or vinyl and wherein the bond shown forms said covalent linkage; and the pharmaceutically acceptable salts thereof; with the proviso that the percentage of ester linkages in said composition are less than 10% of the linkages in the composition as shown by HPLC in conjunction with base hydrolysis.
The resulting porfimer sodium compositions of the present invention are characterized, as stated, in that they contain less than 10% of their oligomeric linkages as ester bonds as shown by base hydrolysis on HPLC assay. These compositions are prepared by employing processes which allow the ester oligomeric linkages to hydrolyze in the reaction process of the acetylated hematoporphyrin composition with alkali, leading to a higher percentage of ether linkages in the product. The ester content of porfimer sodium can be characterized by utilizing HPLC in conjunction with exhaustive base hydrolysis. In this procedure, the product is assayed before and after treatment with base which hydrolyzes the ester bonds. As stated, the product exhibits enhanced stability as compared to the composition of the prior art.
The present invention can therefore be described by the processes by which the porfimer sodium oligomer products are formed. Such B
a process includes the step of reacting the acetylated hematoporphyrin composition with alkali for sufficient time and B
temperature to hydrolyze the e~ter lin~age~ of the oligomeric product~ formed such that there are le~
than 10% by weight ester-lin~ed oligomer~ in the composition. Alternatively, the acetylated hematopor-phyrin composition can be reacted with alk~li,preferably ~odium hydroxide, and the re~ulting product i~ held at an al~aline pH for sufficient timo to hydrolyze the ester lin~age~.
In a preferred embodiment, the prep~ration lo proce~ comprise~ the re~ction of the acetylated hematoporphyrin composition with O.lN-0.2N ~odium hydroxide at room temperature for 16-2~ hour~. The pH
of the ~olution i~ adjusted to 9.~-9.6 with hydrochloric acid and purified by diafiltration with a membrane filter to eliminate low molecular weight components.
In additional aspects, this in~ention provides compositions cont~;ning the predominantly ether lin~ed oligomer composition~ and their use in photodynamic therapy and diagnosis.
--lO- 2094974 DETAI~ED DE8CRIPTION
BRIEF DE8CRIPTION OF TB DRA~ING~
In the specification and claims, reference will be made to the accompanying drawings in which:
FIG. 1 is a graph showing the stability of the compositions of the present invention over time at 2-4C as compared to the composition of the art.
FIG. 2 is a graph showing the stability, over time at 4C, of compositions prepared by reacting the acetylated hematoporphyrin composition with al~ali for a period of 24 hours as compared with a composition of the art.
DE8CRIPTION OF T B PREFERRED EMBODI~NT8 The ~tarting material employed in tho present invention is hematoporphyrin dihy~rochloride.
Hematoporphyrin dihydrochloride is derived from tho naturally occurring compound, heme, and i9 available commercially from for example, Aldrich Chemical Company.
The material i8 availablo in its protonated ~Cl salt form as a dry powder.
2s In accordance with tho pres~nt invention, the hematoporphyrin dihydrochloride starting matorial is reacted with acetic acid/sulfuric acid a~ taught by Lipson et al., J. Natl. Cancer Inst. 26: 1-8 (1961) and Dougherty et al., U.~. Patent 4,649,151 to produce the acetylated hematoporphyrin composition. This compo~ition is then reacted with al~ali, preferably 0.lN-0.2N sodium hydroxide, for ~ufficient ti~o and at sufficient temperature to hydrolyze tho ester lin~ages ~uch that there is less than 10%, proferably lo~s than 4-5% esters in the composition.
In one embodiment, the acetylated hematopor-phyrin i~ reacted with O.lN ~odium hydroxide for at le~st ~n hour at room temperature. The solution is purified using room temperature diafiltration and S stored at 4C from 14 to 21 day~. The m~terial is then repurified u~ing a diafiltration procedure.
In an alternate procedure, the acetylated hematoporphyrin is dissolved in either O.lN or 0.2N
~odium hydroxide and allowed to react for lC to 24 hours at room temperature. The solution is then purified using a room temperature diafiltration procedure or using a diafiltration procedure at 35-40 C.
U~ing an additional method, the acetylated hematoporphyrin is dissolved in either O.lN or 0.2N
sodium hydroxide and allowed to react for one hour at 35-45C. The solution is then purified at room temperature or at 35-45C using a diafiltration procedure.
Alternatively, the acetylated hematoporphyrin i~ di~olved in O.lN sodium hydroxide and allowed to react for one hour at room temperature. Additional sodium hydroxide is then added to a concentration of 0.2N and held there for 10-24 hours or a lN concentra-tion and held for 2 hours. The ~olution is then purified at room temperature using the diafiltration procedure.
In an alternative procedure, acetylated hematoporphyrin i~ di~solved in O.lN sodium hydroxide and allowed to react for one hour at room temperature.
The pH of the ~olution is adjusted to 9.6 with hydrochloric acid and the resulting mixture held for 3 days at room temperature.
The improvement~ over the Dougherty proces~
in each of the aforementioned variation~ re~ult~ in final solutions that are more stable, thu~ allowing -12- 20~4974 ~torag- in the liguid ~tate at l-~C over an extended p-riod of time At pr-sent, th- pr-ferr-d embodim-nt for preparing th- ~table porfimer ~odium compo~ition i~ to react th- acetylat-d hematoporphyrin with O lN ~odium hydroxide and allowing th- reaction to occur for 16-2~
hours The ~olution i~ th-n purified by diafiltration at room temperature The ~olution of the ac-tylated hematopor-phyrin compo~ition is an agu-ou~ ~olution The aqueous alkali ~olution i~ generally in a concentration of O lN-0 2N, but concentrations of up to lN may be used Generally, the ratio of acetylated hematoporphyrin to alkali ~olution i8 1: 50 The time and temperature for the reaction are related Higher temperature~ employ ~horter time~
while lower temperature~ reguir- longer period~ of time to hydrolyze the ester~ Th- concentration of alkali i~ al~o related, in that ~tronger ~lkali concentration~
require ~horter r-action times The temperature is generally moderate, fro~ room temperature to temp-rature~ of 40-~5C ~igher temperature~ are not preferred since degradation of the productQ tends to occur at higher temperature~ The time, temp-rature and ~trength of the alkali Qhould be adequate to hydrolyze the ester bond~ such that about 90% or greater of the oligomer~ are ether linked oligomer~
Thi~ conversion can be monitored by ~uch technique~ as exhaustive ba~e hydrolyQis followed by HP~C ~high performance liguid chromatography) analy~i~ Examples of time~ and temperature~ which give the de~ired re~ults are about 4C for 1~-21 day~, roo~ t-~perature for 16-2~ hour~, 35-~5C for on- hour, room t-~perature for one hour, adju~tment of the pH to 9 6 and holding for 3 days, then purification at room temperature -The oligomer product of the present invention can be characterized in terms of its chemicA~l ma~e-up and properties. Although not understood with certainty, it is believed that the produ~At is a mixture of oligomers in which up to eight porphyrin units which are hematoporphyrin, hydroxyethylvinyldeuteroporphyrin and protoporphyrin units are covalently linked together by ether or ester bonds as described on page ~-5 of this application. It is a property of the present invention that no more than 10% of the composition consists of ester linked oligomers.
The product may be characterized by determining the ester content of the composition using HPLC in conjunction with exhaustive ba~e hydrolysis.
In this procedure, the product is assayed on HPLC to establish a baseline, then subjected to exhaustive treatment with base, such as lN sodium hydroxide for 12-18 hours to hydrolyze all of the ester bonds. The resulting product is then assayed once more on XPLC.
By comparing the HPLC chromatogram before and after base hydrolysis one can determine the percentage ester content in the original composition. As stated, it is a property of the claimed composition that it contains no more than 10% ester lin~ed oligomers when analyzed using this procedure.
Without intending to be bound to any particular theory of how the present invention operates, it is now believed by the present inventors that the invention relies on a series of reactions to yield the composition of the invention. In the fir~t reaction, the acetylated hematoporphyrin composition forms oligomeric lin~ages when reacted with al~ali.
Some of the oligomers formed are ether lin~ages and some are esters. If the reaction i~ allowed to continue for appropriate time and under appropriate conditions, the ester lin~ages hydrolyze and the resulting eompositionQ formed is then predominantly ether linked oligomers whieh show enhaneed stability.
A~ stated above, the compo~itions of the pre~ent invention show enhaneed stability. ~able 1 ~how~ a eompari~on of the purity of the produets produeed by the prior art proee~ aiselosed by U.8.
Patent 4,649,151 (~the Dougherty proees~) ver~u~ three of the variation~ whieh inelude (a) keeping the reaetion of the aeetylated hematoporphyrin and O.lN
sodium hydroxide at room temperature for 24 hour followed by diafiltration at room temperature: (b) ~eeping the reaetion of the aeetylated hematoporphyrin and O.lN ~odium hydroxide for 1 hour at ~0-45C; or te) keeping the reaetion of the aeetylated hematoporphyrin -and O.lN sodium hydroxide at room temperature for 1hour, adjusting the pH to 9.6 with lN hydroehloric aeid, holding for 3 days followed by diafiltration at room temperature. AS is evident from looking at the data in Table 1, analysi~ of the resulting produet in eaeh ease ~how~ that over time while being stored at 2-4C the produet of the Dougherty Proee~s degrades from 92% to 86% while the products obtained with each of the various improvements results in produet~ with only minor degradation.
2s Table I: ~ Porfimer Sodium vs. Storage Time at 2C-4C
Storage Time Dougherty 24-Hour High pH 9.6 (Days) Process Hold Temperature 3 Days 0 92.1 90.8 92.9 91.0 1 9~.3 2 90.3 4 89.8 91.0 6 91.6 7 88.8 93.3 8 91.3 9 87.9 11 87.5 14 87.2 90.6 92.8 92.6 18 86.5 89.4 21 92.0 92.0 23 86.0 28 85.8 91.3 92.3 92.0 34 90.2 92.4 49 90.1 Dougherty Process: 0.lN NaOH 1 hour, purification at RT
24-Hour Hold: 0.lN NaOH 24 hour~, purification at RT
High Temperature: 0.1N NaOH 1 hour at 40-45C, purification at 40-45C
pH 9.6, 3-Day Hold: 0.1N NaOH 1 hour, pH to 9.6 w/lN HCl, 3-day hold, purification at RT
~
A~ further ~hown in Figure 1, the product of the Dougherty Proees~ h~ a con~tant lo~ of purity over time while each of the product~ obtained through the enhaneed proceQ~e~ (a), ~b) and (e) above ~how~
enhanced -Qtability at 2-~C.
AQ ~hown graphieally in Figure 2 the product obtained in eaeh of several preparation~ u~ing the preferred proeedure of reaeting the aeetylated hematoporphyrin and O.lN ~odium hydroxiae at room temperature for 24 hour-Q followed by diafiltration at room temperature retainQ produet ~tability over time ver~u~ produet prepared by the Dougherty proee~ Q .
The porfimer ~odium oligomer produet of thi~
invention ean be formulated into pharmaeeutieal preparations for injeetion. Sueh preparationQ can eontain from 0.1 to about 3.0 gramQ per liter of the porfimer -Qodium material in a pharmaeeutieally aeceptable nonpyrogenie aqueou~ injection vehicle.
Typical preparation~ have pH'~ between about 7.0 and about 8.1 and are iQotonic. They al~o are ~ub~tan-tially free of any solid contaminantQ. The eoncentra-tion, pH and ionic ~trength of the preparation~ can be adjusted by removing water by ultrafiltration, by adding liquid~ ~ueh a~ ~terile water for injeetion, ~terile ~aline for injeetion or baeterio~tatic water for injeetion a~ the~e liquid~ are defined in the book Remington~ Pharmaeeutieal Seienee~, 1985, Maek Publishing Company, 1985, by adding pH adju~ting material~ ~ueh a~ U.S.P. sodium hydroxidQ or ~.8.P.
phosphorie aeid, or the like. The pharmaeeutieal preparations are then paekaged under ~terile eondition~
in ~uitable vialQ, bottle~, prefilled ~yringe~, ana the like for admini~tration via intravenou~ infu~ion or intramu~eular injeetion to patient~.
The porfimer Qodium oligomer produet~ of the invention may be u~ed in photodynamic therapy or d~agno~is in the manner which i~ di~clo~ed for such porphyrin based product~. The amount of porfimer ~odium oligomer product administered to the patent can range from about 0.1 mg to 10 mg per ~ilogram of body weight with do~age~ of 1 to 2 mg/~g being preferred.
The invention will now be further de~cribed with reference to the following Example~. The~e are presented ~olely to ma~e clear preferred embodiments of the pre~ent invention and are not to be con~trued a~
limiting the ~cope of the invention which i~ as defined by the appended claim~.
~xample 1 Acetylated hematoporphyrin (1 part by weight) i5 dissolved in O.lN ~odium hydroxide (50 part~ by volume) and stirred for one hour at room temperature.
After the ~tir period the ~olution i~ adju~ted to pH
9.4 to 9.6 with lN hydrochloric acid. It i8 filtered through a 5~m filter and then concentrated to 1/4(12.5 part~) of its original volume in an ultrafilter with 10,000 molecular weight cut off membrane~. The solution i~ then purified via diafiltration maint~;~ing con~tant volume with 120 volume~ of water and ~eeping the pH at 9.~ to 9.6 with O.lN ~odium hydroxide. Thi~
i~ also done at room temperature. After the purification, the ~olution i~ removed from the ultrafilter, diluted to 3/8(18.8 parts) of it~ original volume and pH adju~ted to 7.5 to 7.7 with lN
hydrochloric acid. The ~olution i~ then ~tored at 4C
for 14 to 21 day~. After ~torage, the ~olution i~ pH
adju~ted to 9.4 to 9.6 with O.lN ~odium hydroxide and concentrated to 1/4(12.5 part~) of it~ original volume.
The solution i8 then repurified a~ above. The ~olution iq diluted to 3/8(18.8 part~) of it~ original volume and pH adju~ted to 7.5 to 7.7 with lN hydrochloric acid. The solution i~ then analyzed and, if nece~ary, it i~ adjusted to between 13 to 18 mg/mL by the addition of water The solution i~ then filtered through a 0 22~m filter into bottles for storage at 1C
to 4C to await further processing ~xample 2 Acetylated h-matoporphyrin ~1 part by weight) is dissolved in O lN or 0 2N sodium hydroxid- ~50 parts by volume) and stirr-d for 16 to 24 hours at room te~perature After th- stir period th- solution is adjusted to p~ 9 4 to 9 6 with lN hydrochloric acid It is filtered through a 5~ filter and th-n concentrated to 1/4~12 5 part~) of its original volume in an ultrafilter with 10,000 molecuiar w-ight cut off membranes The solution i~ then purifi-d via diafiltration at room temperature or at 35C to 45C, maintaining constant volume with 120 volumes of water and keeping the p~ at 9 4 to 9 6 with O lN sodium hydroxide After tho purification, th- solution is removed from the ultrafiltor, diluted to 3/8(18 8 part~) of it~ original ~olu~e and pH adjusted to 7 5 to 7 7 with lN hydrochloric acid The solution is then analyzed and, if necessary, it is ad~ust-d to between 13 to 18 mg/m~ by the addition of ~ater Th- ~olution is then filt-red through a 0 22~m filter into bottles for storage at 1C to 4C to a~ait further processing ~xamDl- 3 Acetylated hematoporphyrin (1 part by weight) is dissolved in O lN or 0 2N ~odiu~ hydroxid- (50 parts by volum-) and stirr-d for one hour at 3S to 45C
After the stir period the solution i~ ad~ust-d to pH
9 4 to 9 6 with lN hydrochloric aci~ It i~ filter-d through a 5~m filt-r and then conc-ntratsd to 1/~(12 5 parts) of its original volume in an ultrafilter with 10,000 molecular weight cut off me~bran-s Th-solution is then purified via diafiltration at room temperature or at 35 to 45C, maint-in~ng constant volume with 120 volum-s of water and ~eeping the p~ at 9.4 to 9.6 with o.lN sodium hydroxide. After the purification, the _olution is removed from the ultrafilter, diluted to 3/8(18.8 part_) of itQ original volum- and pH adjusted to 7.5 to 7.7 with lN
s hydrochlori¢ acid. The solution is then analyzed and, if nece_sary, it is adjuQted to between 13 to 18 mg/mL
by the addition of water. The solution is then filtered through a 0.22~m filter into bottleQ for storage at 1C to 4C to await further processing.
~xample 4 Acetylated hematoporphyrin ~1 P~rt by weight) i~ di~Qolved in 0.lN sodium hydro~ide ~50 parts by volumo) and stirred for 1 hour at room temperature.
Sodium hydroxide is then added to give an overall concentration ~based on initial NaO~ plus this addition) of either l.lN ~2 parts NaOH) or 0.2N ~0.2 parts NaOH). ThiQ solution is then stirred for 2 hours (l.lN NaOH) or 24 hours ~0.2N NaOH). After the stir period the solution is ad~usted to pH 9.4 to 9.6 with lN hydrochloric acid. It is th-n filtered through a 5~m filter and then conc-ntrated to 1/4~12.5 parts) of it~ original volume in an ultrafilter with 10,000 molecular weight cut off membranes. The solution i_ then purified via diafiltration maintaining constant volume with 120 volume~ of water and keeping the p~ at 9.4 to 9.6 with 0.lN sodium hydro~id-. This is also done at roo~ temperature. Aft-r the purification, the solution is removed from th- ultrafilt-r, diluted to 3/8~18.8 parts) of its original volume and p~ adjusted to 7.5 to 7.7 with lN hydrochloric acid. The solution is then analyzed and, if n-c-ssary, it is ad~usted to between 13 to 18 mg/~L by tho addition of water. The solution is th-n filtered through a 0.22~m filter into bottles for storage at ~C to 4C to await further proce~sing.
~xample 5 Acetylated hematoporphyrin (1 part by weight) is dissolved in O.lN sodium hydroxide (50 part~ by volume) and stirred for one hour at room temperature.
After the stir period the ~olution i8 adju~ted to pH
9.4 to 9.6 with lN hydrochloric acid and i~ held at room temperature for 3 day~. After the holding period, the solution i~ filtered through a S~m filter and then concentrated to 1/4(12.5 part~) of it~ original volume in an ultrafilter with 10,000 molecular weight cut off membrane~. The solution i~ then purified via diafiltration maintaining con~tant volume with 120 volume~ of water and ~eeping the pH at 9.4 to 9.6 with o.lN sodium hydroxide. Thi~ i~ al~o done at room temperature. After the purification, the solution i~
removed from the ultrafilter, diluted to 3/8~18.8 parts) of it~ original volume and pH adju~ted to 7.5 to 7.7 with lN hydrochloric acid. The ~olution i~ then analyzed and, if necessary, it is adju~ted to between 13 to 18 mg/mL by the addition of water. The ~olution is then filtered through a 0.22~m filter into bottle~
for storage at 1C to 4C to await further proce~ing.
Claims (8)
1. A porfimer sodium pharmaceutical composition comprised of porphyrin oligomers having ether and ester covalent linkages wherein less than 10% of said linkages in the porphyrin oligomers are ester linkages, in association with a pharmaceutically acceptable carrier.
2. A biologically active composition being fluorescent, photosensitizing and having the capability of localizing in and being retained in tumor tissue as compared to normal tissues which composition comprises a mixture of porphyrin oligomers, said oligomers comprised of two to eight porphyrin molecules covalently linked by ether and ester bonds wherein at least one porphyrin molecule has the formula:
wherein R is hyroxyethyl or vinyl and wherein the bond shown forms said covalent linkage, and the pharmaceutically acceptable salts thereof; with the proviso that the percentage of ester linkages in said composition are less than 10% of the linkages in the composition.
wherein R is hyroxyethyl or vinyl and wherein the bond shown forms said covalent linkage, and the pharmaceutically acceptable salts thereof; with the proviso that the percentage of ester linkages in said composition are less than 10% of the linkages in the composition.
3. A process for the preparation of a porfimer sodium pharmaceutical composition comprises of porphyrin oligomers having ether and ester linkages which comprises treating acetylated hematoporphyrin with alkali for sufficient time and sufficient temperature to reduce the amount of porphyrin oligomers joined by ester linkages to less than 10% of the linkages in the composition.
4. A process according to claim 3 wherein the alkali is sodium hydroxide.
5. In a process for the preparation of a porfimer sodium composition by reaction of acetylated hematoporphyrin with sodium hydroxide, the improvement which comprises contacting the acetylated hematoporphyrin with 0.1N sodium hydroxide for at least one hour, storing the solution at 4° for 14 to 21 days and purifying by diafiltration.
6. In a process for the preparation of a porfimer sodium by reaction of acetylated hematoporphyrin with sodium hydroxide, the improvement which comprises contacting the acetylated hematoporphyrin with at least 0.1N sodium hydroxide for 16 to 24 hours at a temperature less than 45°C and purifying by diafiltration at a temperature less than 45°C.
7. In a process for the preparation of a porfimer sodium by reaction of acetylated hematoporphyrin with sodium hydroxide, the improvement which comprises contacting the acetylated hematoporphyrin with at least 0.1N sodium hydroxide for at least one hour, increasing the concentration to less than 1N but at least 0.2N for 2 to 24 hours, and purifying by diafiltration at room temperature.
8. A pharmaceutical composition prepared by the process of claims 3, 4, 5, 6 or 7.
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US873,860 | 1992-04-27 | ||
US07/873,860 US5244914A (en) | 1992-04-27 | 1992-04-27 | Stable porfimer sodium compositions and methods for their manufacture |
Publications (2)
Publication Number | Publication Date |
---|---|
CA2094974A1 CA2094974A1 (en) | 1993-10-28 |
CA2094974C true CA2094974C (en) | 1996-10-22 |
Family
ID=25362473
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CA002094974A Expired - Lifetime CA2094974C (en) | 1992-04-27 | 1993-04-27 | Stable porfimer sodium compositions and methods for their manufacture |
Country Status (10)
Country | Link |
---|---|
US (2) | US5244914A (en) |
EP (1) | EP0568323B1 (en) |
JP (1) | JP2928047B2 (en) |
KR (1) | KR100221206B1 (en) |
AT (1) | ATE150968T1 (en) |
CA (1) | CA2094974C (en) |
DE (1) | DE69309340T2 (en) |
DK (1) | DK0568323T3 (en) |
ES (1) | ES2104056T3 (en) |
GR (1) | GR3023978T3 (en) |
Families Citing this family (67)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US5371199B1 (en) * | 1992-08-14 | 1995-12-26 | Univ Pennsylvania | Substituted porphyrins porphyrin-containing polymers and synthetic methods therefor |
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US9371555B2 (en) | 2012-06-01 | 2016-06-21 | Concordia Laboratories Inc. | Lighting systems and methods of using lighting systems for in vitro potency assay for photofrin |
AU2014216112B2 (en) | 2013-02-15 | 2019-02-21 | Allergan, Inc. | Sustained drug delivery implant |
MA47786A (en) | 2017-03-14 | 2020-01-22 | Ohio State Innovation Foundation | METHODS AND COMPOSITIONS RELATING TO IGG3 IMMUNOCONJUGATES TARGETING TISSUE FACTOR |
KR102182630B1 (en) | 2019-12-19 | 2020-11-24 | 김진왕 | Eco-friendly smart photosensitizers and photobiomodulation of stem cell therapy agent using the same |
Family Cites Families (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4649151A (en) * | 1982-09-27 | 1987-03-10 | Health Research, Inc. | Drugs comprising porphyrins |
US4882234A (en) * | 1986-11-12 | 1989-11-21 | Healux, Inc. | Storage-stable porphin compositions and a method for their manufacture |
US4861876A (en) * | 1986-11-26 | 1989-08-29 | Wayne State University | Hematoporphyrin derivative and method of preparation and purification |
US5059619A (en) * | 1989-06-14 | 1991-10-22 | Quadra Logic Technologies, Inc. | Stable freeze-dried polyhematoporphyrin ether/ester |
-
1992
- 1992-04-27 US US07/873,860 patent/US5244914A/en not_active Expired - Lifetime
-
1993
- 1993-04-27 JP JP5101601A patent/JP2928047B2/en not_active Expired - Lifetime
- 1993-04-27 KR KR1019930007074A patent/KR100221206B1/en not_active IP Right Cessation
- 1993-04-27 CA CA002094974A patent/CA2094974C/en not_active Expired - Lifetime
- 1993-04-27 EP EP93303290A patent/EP0568323B1/en not_active Expired - Lifetime
- 1993-04-27 DK DK93303290.6T patent/DK0568323T3/en active
- 1993-04-27 ES ES93303290T patent/ES2104056T3/en not_active Expired - Lifetime
- 1993-04-27 DE DE69309340T patent/DE69309340T2/en not_active Expired - Lifetime
- 1993-04-27 AT AT93303290T patent/ATE150968T1/en active
- 1993-06-10 US US08/075,227 patent/US5438071A/en not_active Expired - Lifetime
-
1997
- 1997-07-02 GR GR970401631T patent/GR3023978T3/en unknown
Also Published As
Publication number | Publication date |
---|---|
DE69309340D1 (en) | 1997-05-07 |
KR930021143A (en) | 1993-11-22 |
KR100221206B1 (en) | 1999-10-01 |
GR3023978T3 (en) | 1997-10-31 |
ES2104056T3 (en) | 1997-10-01 |
EP0568323B1 (en) | 1997-04-02 |
JP2928047B2 (en) | 1999-07-28 |
DE69309340T2 (en) | 1997-07-17 |
DK0568323T3 (en) | 1997-07-07 |
ATE150968T1 (en) | 1997-04-15 |
EP0568323A1 (en) | 1993-11-03 |
CA2094974A1 (en) | 1993-10-28 |
US5244914A (en) | 1993-09-14 |
US5438071A (en) | 1995-08-01 |
JPH06220062A (en) | 1994-08-09 |
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