CA2421485A1 - Water soluble rapamycin esters - Google Patents
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- CA2421485A1 CA2421485A1 CA002421485A CA2421485A CA2421485A1 CA 2421485 A1 CA2421485 A1 CA 2421485A1 CA 002421485 A CA002421485 A CA 002421485A CA 2421485 A CA2421485 A CA 2421485A CA 2421485 A1 CA2421485 A1 CA 2421485A1
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D498/00—Heterocyclic compounds containing in the condensed system at least one hetero ring having nitrogen and oxygen atoms as the only ring hetero atoms
- C07D498/12—Heterocyclic compounds containing in the condensed system at least one hetero ring having nitrogen and oxygen atoms as the only ring hetero atoms in which the condensed system contains three hetero rings
- C07D498/18—Bridged systems
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
- A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
- A61K47/56—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule
- A61K47/59—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyureas or polyurethanes
- A61K47/60—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyureas or polyurethanes the organic macromolecular compound being a polyoxyalkylene oligomer, polymer or dendrimer, e.g. PEG, PPG, PEO or polyglycerol
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P11/00—Drugs for disorders of the respiratory system
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- A61P19/00—Drugs for skeletal disorders
- A61P19/02—Drugs for skeletal disorders for joint disorders, e.g. arthritis, arthrosis
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P25/00—Drugs for disorders of the nervous system
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/10—Antimycotics
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P37/00—Drugs for immunological or allergic disorders
- A61P37/02—Immunomodulators
- A61P37/06—Immunosuppressants, e.g. drugs for graft rejection
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P9/00—Drugs for disorders of the cardiovascular system
Abstract
This invention provide pegylated hydroxyesters of rapamycin which are useful in inducing immunosuppression and in the treatment of transplantation rejection, autoimmune diseases, solid tumors, fungal infections, and vascula r disease.
Description
WATER SOLUBLE RAPAMYCIN ESTERS
This invention relates to water soluble rapamycin esters which are useful in inducing immunosuppression and in the treatment of transplantation rejection, autoimmune diseases, solid tumors, fungal infections, and vascular disease.
More particularly, this invention concerns methoxypoly(ethylene glycol) esters of hydroxy-esters of rapamycin, their preparation and intermediates therein and methods for using them for inducing immunosuppression, and in the treatment of transplantation rejection, graft vs. host disease, autoimmune diseases, diseases of inflammation, adult T-cell leukemiahymphoma, solid tumors, fungal infections, cardiovascular disease, cerebral vascular disease, peripheral vascular disease or hyperproliferative vascular disorders.
Rapamycin is a macrocyclic triene antibiotic produced by Streptom~es h~rgroscopicus, which was found to have antifungal activity, particularly against Candida albicans, both in vitro and in vivo [C. Vezina et al., J. Antibiot. 28, 721 (1975); S.N.
Sehgal et al., J. Antibiot. 28, 727 (1975); H. A. Baker et al., J. Antibiot.
31, 539 (1978);
U.S. Patent 3,929,992; and U.S. Patent 3,993,749]. Additionally, rapamycin alone (U.S.
Patent 4,885,171) or in combination with picibanil (U.S. Patent 4,401,653) has been shown to have antitumor activity.
The immunosuppressive effects of rapamycin have been disclosed in FASEB 3, 3411 (1989). Cyclosporin A and FK-506, other macrocyclic molecules, also have been shown to be effective as immunosuppressive agents, therefore useful in preventing transplant rejection [FASEB 3, 3411 (1989); FASEB 3, 5256 (1989); R. Y. Calne et al., Lancet 1183 (1978); and U.S. Patent 5,100,899]. R. Martel et al. [Can. J.
Physiol.
Pharmacol. 55, 48 (1977)] disclosed that rapamycin is effective in the experimental allergic encephalomyelitis model, a model for multiple sclerosis; in the adjuvant arthritis model, a model for rheumatoid arthritis; and effectively inhibited the formation of IgE-like antibodies.
Rapamycin is also useful in preventing or treating systemic lupus erythematosus [U.S. Patent 5,078,999], pulmonary inflammation [U.S. Patent 5,080,899], insulin dependent diabetes mellitus [U.S. Patent 5,321,009], skin disorders, such as psoriasis [U.S. Patent 5,286,730], bowel disorders [U.S. Patent 5,286,731], smooth muscle cell proliferation and intimal thickening following vascular injury [U.S. Patents 5,288,711 and 5,516,781], adult T-cell leukemia/lymphoma [European Patent Application 525,960 A1], ocular inflammation [U.S. Patent 5,387,589], malignant carcinomas [U.S. Patent 5,206,018], cardiac inflammatory disease [U.S. Patent 5,496,832], and anemia [U.S.
Patent 5,561,138].
A rapamycin ester, rapamycin 42-ester with 3-hydroxy-2-(hydroxymethyl)-2 methylpropionic acid [disclosed in U.S. Patent 5,362,718], also known as CCI-779, has been shown to have antitumor activity against a variety of tumor cell lines, in in vivo animal tumor models, and in Phase I clinical trials. [Gibbons, J., Proc. Am:
Assoc. Can.
Res. 40: 301 (1999); Geoerger, B., Proc. Am. Assoc. Can. Res. 40: 603 (1999);
Alexandre, J., Proc. Am. Assoc. Can. Res. 40: 613 (1999); and Alexandre, J., Clin.
Cancer. Res. 5 (November Supp.): Abstr. 7 (1999)].
Polyethylene glycol (PEG) is a linear or branched, neutral polymer available in a variety of molecular weights and is soluble in water and most organic solvents. At molecular weights less than 1000 are the viscous, colorless liquids; higher molecular weight PEGs are waxy, white solids. The melting point of the solid is proportional to the molecular weight, approaching a plateau at 67°C. Molecular weight range from a few hundred to approximately 20,000 are commonly used in biological and biotechnological applications. Of much interest in the biomedical areas is the fact that PEG is nontoxic and was approved by FDA for internal consumption. Pegylated rapamycin is disclosed in US Patent 5,780,462.
DESCRIPTION OF THE INVENTION
This invention provides methoxypoly(ethylene glycol) esters of hydroxyesters of rapamycin having the structure ~nl (A) wherein R1 and R2 are each, independently, hydrogen or -CO(CR3R4)b(CR5R6)dCR~R$R9 ;
R3 and R4 are each, independently, hydrogen, alkyl of 1-6 carbon atoms, alkenyl of 2-7 carbon atoms, alkynyl of 2-7 carbon atoms, trifluoromethyl, or -F;
R5 and R6 are each, independently, hydrogen, alkyl of 1-6 carbon atoms, alkenyl of 2-7 carbon atoms, alkynyl of 2-7 carbon atoms, -(CR3R4)fORlo, -CF3, -F, or -C02R11;
R~ is hydrogen, alkyl of 1-6 carbon atoms, alkenyl of 2-7 carbon atoms, alkynyl of 2-7 carbon atoms, -(CR3R4)fORI~, -CF3, -F, or-CO~R11;
R$ and R9 are each, independently, hydrogen, alkyl of 1-6 carbon atoms, alkenyl of 2-7 carbon atoms, alkynyl of 2-7 carbon atoms, -(CR3R4)fORI~, -CFg, -F, or -C02R11;
R1~ is hydrogen or-COCH2-S-CH2CH2-(O-CH2-CH2)n-OCHg;
R11 is hydrogen, alkyl of 1-6 carbon atoms, alkenyl of 2-7 carbon atoms, alkynyl of 2-7 carbon atoms, or phenylalkyl of 7-10 carbon atoms;
b = 0-6;
d = 0-6;
f = 0-6;
n = 5-450;
with the proviso that R1 and R2 are both not hydrogen and further provided that either R1 or R2 contains at least one -(CR3R~)fORI~ group in which R1~ is -COCH2-S-CH2CH2-(O-CH2-CH2)n-OCHg, or a pharmaceutically acceptable salt thereof which are useful for inducing immunosuppression, and in the treatment of transplantation rejection, graft vs. host disease, autoimmune diseases, diseases of inflammation, adult T-cell leukemia/lymphoma, solid tumors, fungal infections, cardiovascular disease, cerebral vascular disease, peripheral vascular disease or hyperproliferative vascular disorders. The compounds of this invention can also be referred to as pegylated hydroxyesters of rapamycin.
In the compounds (A) and (B) of this invention the following are examples of variables taken singly, in any combination or all:
R2 may be for example hydrogen.
This invention relates to water soluble rapamycin esters which are useful in inducing immunosuppression and in the treatment of transplantation rejection, autoimmune diseases, solid tumors, fungal infections, and vascular disease.
More particularly, this invention concerns methoxypoly(ethylene glycol) esters of hydroxy-esters of rapamycin, their preparation and intermediates therein and methods for using them for inducing immunosuppression, and in the treatment of transplantation rejection, graft vs. host disease, autoimmune diseases, diseases of inflammation, adult T-cell leukemiahymphoma, solid tumors, fungal infections, cardiovascular disease, cerebral vascular disease, peripheral vascular disease or hyperproliferative vascular disorders.
Rapamycin is a macrocyclic triene antibiotic produced by Streptom~es h~rgroscopicus, which was found to have antifungal activity, particularly against Candida albicans, both in vitro and in vivo [C. Vezina et al., J. Antibiot. 28, 721 (1975); S.N.
Sehgal et al., J. Antibiot. 28, 727 (1975); H. A. Baker et al., J. Antibiot.
31, 539 (1978);
U.S. Patent 3,929,992; and U.S. Patent 3,993,749]. Additionally, rapamycin alone (U.S.
Patent 4,885,171) or in combination with picibanil (U.S. Patent 4,401,653) has been shown to have antitumor activity.
The immunosuppressive effects of rapamycin have been disclosed in FASEB 3, 3411 (1989). Cyclosporin A and FK-506, other macrocyclic molecules, also have been shown to be effective as immunosuppressive agents, therefore useful in preventing transplant rejection [FASEB 3, 3411 (1989); FASEB 3, 5256 (1989); R. Y. Calne et al., Lancet 1183 (1978); and U.S. Patent 5,100,899]. R. Martel et al. [Can. J.
Physiol.
Pharmacol. 55, 48 (1977)] disclosed that rapamycin is effective in the experimental allergic encephalomyelitis model, a model for multiple sclerosis; in the adjuvant arthritis model, a model for rheumatoid arthritis; and effectively inhibited the formation of IgE-like antibodies.
Rapamycin is also useful in preventing or treating systemic lupus erythematosus [U.S. Patent 5,078,999], pulmonary inflammation [U.S. Patent 5,080,899], insulin dependent diabetes mellitus [U.S. Patent 5,321,009], skin disorders, such as psoriasis [U.S. Patent 5,286,730], bowel disorders [U.S. Patent 5,286,731], smooth muscle cell proliferation and intimal thickening following vascular injury [U.S. Patents 5,288,711 and 5,516,781], adult T-cell leukemia/lymphoma [European Patent Application 525,960 A1], ocular inflammation [U.S. Patent 5,387,589], malignant carcinomas [U.S. Patent 5,206,018], cardiac inflammatory disease [U.S. Patent 5,496,832], and anemia [U.S.
Patent 5,561,138].
A rapamycin ester, rapamycin 42-ester with 3-hydroxy-2-(hydroxymethyl)-2 methylpropionic acid [disclosed in U.S. Patent 5,362,718], also known as CCI-779, has been shown to have antitumor activity against a variety of tumor cell lines, in in vivo animal tumor models, and in Phase I clinical trials. [Gibbons, J., Proc. Am:
Assoc. Can.
Res. 40: 301 (1999); Geoerger, B., Proc. Am. Assoc. Can. Res. 40: 603 (1999);
Alexandre, J., Proc. Am. Assoc. Can. Res. 40: 613 (1999); and Alexandre, J., Clin.
Cancer. Res. 5 (November Supp.): Abstr. 7 (1999)].
Polyethylene glycol (PEG) is a linear or branched, neutral polymer available in a variety of molecular weights and is soluble in water and most organic solvents. At molecular weights less than 1000 are the viscous, colorless liquids; higher molecular weight PEGs are waxy, white solids. The melting point of the solid is proportional to the molecular weight, approaching a plateau at 67°C. Molecular weight range from a few hundred to approximately 20,000 are commonly used in biological and biotechnological applications. Of much interest in the biomedical areas is the fact that PEG is nontoxic and was approved by FDA for internal consumption. Pegylated rapamycin is disclosed in US Patent 5,780,462.
DESCRIPTION OF THE INVENTION
This invention provides methoxypoly(ethylene glycol) esters of hydroxyesters of rapamycin having the structure ~nl (A) wherein R1 and R2 are each, independently, hydrogen or -CO(CR3R4)b(CR5R6)dCR~R$R9 ;
R3 and R4 are each, independently, hydrogen, alkyl of 1-6 carbon atoms, alkenyl of 2-7 carbon atoms, alkynyl of 2-7 carbon atoms, trifluoromethyl, or -F;
R5 and R6 are each, independently, hydrogen, alkyl of 1-6 carbon atoms, alkenyl of 2-7 carbon atoms, alkynyl of 2-7 carbon atoms, -(CR3R4)fORlo, -CF3, -F, or -C02R11;
R~ is hydrogen, alkyl of 1-6 carbon atoms, alkenyl of 2-7 carbon atoms, alkynyl of 2-7 carbon atoms, -(CR3R4)fORI~, -CF3, -F, or-CO~R11;
R$ and R9 are each, independently, hydrogen, alkyl of 1-6 carbon atoms, alkenyl of 2-7 carbon atoms, alkynyl of 2-7 carbon atoms, -(CR3R4)fORI~, -CFg, -F, or -C02R11;
R1~ is hydrogen or-COCH2-S-CH2CH2-(O-CH2-CH2)n-OCHg;
R11 is hydrogen, alkyl of 1-6 carbon atoms, alkenyl of 2-7 carbon atoms, alkynyl of 2-7 carbon atoms, or phenylalkyl of 7-10 carbon atoms;
b = 0-6;
d = 0-6;
f = 0-6;
n = 5-450;
with the proviso that R1 and R2 are both not hydrogen and further provided that either R1 or R2 contains at least one -(CR3R~)fORI~ group in which R1~ is -COCH2-S-CH2CH2-(O-CH2-CH2)n-OCHg, or a pharmaceutically acceptable salt thereof which are useful for inducing immunosuppression, and in the treatment of transplantation rejection, graft vs. host disease, autoimmune diseases, diseases of inflammation, adult T-cell leukemia/lymphoma, solid tumors, fungal infections, cardiovascular disease, cerebral vascular disease, peripheral vascular disease or hyperproliferative vascular disorders. The compounds of this invention can also be referred to as pegylated hydroxyesters of rapamycin.
In the compounds (A) and (B) of this invention the following are examples of variables taken singly, in any combination or all:
R2 may be for example hydrogen.
b and d for example may be independently 0 or 1; e.g. b is 0 and d is 1;
preferably both are 0.
Examples of f are 1-4 , e.g. 1-2, preferably f is 1.
R3 and R4 may be hydrogen or alkyl of 1-6 carbon atoms, preferably hydrogen.
R~ may for example be hydrogen, alkyl of 1-6 carbon atoms, alkenyl of 2-7 carbon atoms, alkynyl of 2-7 carbon atoms or -CFg; e.g. hydrogen~or alkyl of 1-4 carbon atoms.
R$ and R9 may be for example be hydrogen or-(CR3R4)fORlO
Where more than one R1~ _ -COCH2-S-CH2CH2-(O-CH2-CH2)n-OCHg group is present then they may be the same or different. Preferably there are one or two such groups present.
Examples of n range from 5-400 such as 5-350 or 5-300 e.g. in the range from about 65-155.
When- applicable, pharmaceutically acceptable salts can be formed from organic and inorganic bases (i.e., when a compound contains a free hydroxyl group), such as alkali metal salts (for example, sodium, lithium, or potassium) alkaline earth metal salts, ammonium salts, alkylammonium salts containing 1-6 carbon atoms or dialkylammonium salts containing 1-6 carbon atoms in each alkyl group, and trialkylammonium salts containing 1-6 carbon atoms in each alkyl group, when the rapamycin contains a suitable acidic moiety.
As used in accordance with this invention, the term "providing" with respect to providing a compound or substance covered by this invention, means either directly administering such a compound or substance, or administering a prodrug, derivative, or analog which will form an effective amount of the compound or substance within the body.
Of the pegylated hydroxyesters of rapamycin covered by this invention, it is preferred that the hydroxyester of rapamycin is CCI-779, in which one or both of the hydroxyl groups of the 3-hydroxy-2-(hydroxymethyl)-2-methylpropionic acid moiety are pegylated. Of the compounds of this invention, it is preferred that n = 5 -200; more preferred that n = 8 - 135. Most preferred members are those in which n = 8 -20 and those in which n = 90 - 120. The values of n refer to the range of repeating ethoxy units in the PEG side chain. For example, when a compound is described as having n =
preferably both are 0.
Examples of f are 1-4 , e.g. 1-2, preferably f is 1.
R3 and R4 may be hydrogen or alkyl of 1-6 carbon atoms, preferably hydrogen.
R~ may for example be hydrogen, alkyl of 1-6 carbon atoms, alkenyl of 2-7 carbon atoms, alkynyl of 2-7 carbon atoms or -CFg; e.g. hydrogen~or alkyl of 1-4 carbon atoms.
R$ and R9 may be for example be hydrogen or-(CR3R4)fORlO
Where more than one R1~ _ -COCH2-S-CH2CH2-(O-CH2-CH2)n-OCHg group is present then they may be the same or different. Preferably there are one or two such groups present.
Examples of n range from 5-400 such as 5-350 or 5-300 e.g. in the range from about 65-155.
When- applicable, pharmaceutically acceptable salts can be formed from organic and inorganic bases (i.e., when a compound contains a free hydroxyl group), such as alkali metal salts (for example, sodium, lithium, or potassium) alkaline earth metal salts, ammonium salts, alkylammonium salts containing 1-6 carbon atoms or dialkylammonium salts containing 1-6 carbon atoms in each alkyl group, and trialkylammonium salts containing 1-6 carbon atoms in each alkyl group, when the rapamycin contains a suitable acidic moiety.
As used in accordance with this invention, the term "providing" with respect to providing a compound or substance covered by this invention, means either directly administering such a compound or substance, or administering a prodrug, derivative, or analog which will form an effective amount of the compound or substance within the body.
Of the pegylated hydroxyesters of rapamycin covered by this invention, it is preferred that the hydroxyester of rapamycin is CCI-779, in which one or both of the hydroxyl groups of the 3-hydroxy-2-(hydroxymethyl)-2-methylpropionic acid moiety are pegylated. Of the compounds of this invention, it is preferred that n = 5 -200; more preferred that n = 8 - 135. Most preferred members are those in which n = 8 -20 and those in which n = 90 - 120. The values of n refer to the range of repeating ethoxy units in the PEG side chain. For example, when a compound is described as having n =
200, it means that such compound consists of a mixture of compounds having a normal distribution between n = 5 and n = 200, with approximately n = 100 having the greatest frequency. With compounds III and IV, the average n was 108, and 99°l0 of n being between 65 and 155. The compounds of this invention may also be described and understood based upon the average molecular weight of the polyethylene glycol chains used to produce their ester chains. For instance, an CCI-779-PEG 5000 ester refers to a compound of the general formula above in which one side chain PEG ester is formed utilizing a polyethylene glycol derivative having an average molecular weight range at or near 5,000; and CCI-779-(PEG 5000)2 ester refers to a compound of the general formula above in two side chain PEG esters are formed utilizing a polyethylene glycol derivative having an average molecular weight range at or near 5,000.
The terms "alkyl", "alkenyl" and "alkynyl" include both straight chain as well as branched carbon chains. Alkyl groups may contain 1-6, e.g. 1-4 carbon atoms, eg methyl, ethyl, propyl and tert-butyl. Alkenyl or alkynyl may contain 2-7 carbon atoms eg 2- 5 such as vinyl, propenyl or acetylenyl.
This invention also provides a process for preparing compounds of formula (A) which comprises reacting a compound of formula (B):
ORA
~'OMe ., O O ~ ORB
3i O O O Me0'' O
HO
'O OMe '~ ~ / ~ /
I ' wherein RA is R1 as hereinbefore defined, and wherein RB is R2 as hereinbefore defined, and in which R10 is as hereinbefore defined in connection with formula (A) providing that at least one R10 group represents -COCH2-X where X is a suitable leaving group;
with a compound of formula (C):
HS-CH2CH2-(O-CH2-CH2)n-OCH3 (C) wherein n is as defined above, if necessary any reactive substituent group being protected and removing any protecting group as required, to give a compound of formula (A) or a pharmaceutically acceptable salt thereof.
Compounds of formula (B) also form another aspect of this invention.
The reagents used in the preparation of the compounds of this invention can be either commercially obtained or can be prepared by standard procedures described in the literature.
The preparation of the hydroxyesters of rapamycin, from which the pegylated hydroxyesters are made from, are described in US Patent 5,362,718 and WO
95/28406, which is hereby incorporated by reference. Conveniently one or more of the hydroxyesters may be acylated with a acylating agent having the general structure X-CH2CO~H, where X is a suitable leaving group, such as bromine or iodine, in the presence of a coupling agent such as dicyclohexylcarbodiimide (DCC) in the presence of a base catalyst, such as dimethylaminopyridine (DMAP). Accordingly, rapamycin ester with 3-hydroxy-2-(2-iodo-acetoxymethyl)-2-methyl-propionic acid (Compound I) and rapamycin 42-ester with 3-(2-lodo-acetoxy)-2-(2-iodo-acetoxymethyl)-2-methyl propionic acid (Compound II) are intermediates prepared using this methodology, that are useful in the production of the corresponding pegylated hydroxyesters of rapamycin (Compounds III and IV, respectively).
The pegylated esters may be conveniently be produced utilizing the polyethylene glycols known in the art, such as those described on pages 355 to 361 of the Handbook of Pharmaceutical Excipients, Second Edition, 1994. The preferred compounds of this invention may also be described as those of the formula esterified using polyethylene glycols having an average molecular weight of from about 200 to about 200,000. A preferred range of the PEG esters of this invention includes those in which the molecular weight of the polyethylene glycol portion of the ester chain has a molecular weight in the range of from about 300 to about 20,000, more preferably between about 350 and about 6,000. Reaction of the hydroxyester of rapamycin which has been acylated as described above, with a suitable polyethyleneglycol thiol in the presence of a base, such as sodium bicarbonate provides the desired pegylated hydroxyester of rapamycin.
The antitumor activity of the compounds of this invention were confirmed in a standard pharmacological test procedure which measures the inhibition of U87MG
human glioblastoma cell growth (as a function of 3H-thymidine incorporation), using CCI-779-PEG 5000 ester (Compound III) and CCI-779-(PEG 5000)2 ester (Compound IV) as representative compounds of this invention. The following briefly describes the procedure used and results obtained. U87MG human glioblastoma cells (ATCC #
HTB-14; available from the American Type Culture Collection; 10801 University Boulevard;
Manassas, VA 20110;), were grown in the following media.
Growth Medium: BRL Minimum Essential Medium with Earle Salts (500 mL) + 5 mL BRL MEM Non-Essential Amino Acids (lOmM) +5 mL BRL Penicillin-Streptomycin ( 10000 u/m L,10000 ~g/m L) +5 mL BRL Na Pyruvate Solution (100mM) +5 mL BRL L-Glutamine 200mM
+50 mL BRL Fetal Bovine Serum (Qualified) Test Procedure:
1. Cells were trypsinized and plated at a concentration of 104 cells/well in a final volume of 200 pL growth medium in 96-well flat bottom plates and allowed to adhere for 24 hours at 37°C.
2. The media was removed by aspiration with care to not disturb the cell monolayer.
200 p,L of fresh growth media was added per well, allowing enough wells for samples to be run in triplicate. Test compounds were added in 10 p.L phosphate buffer solution (PBS) and incubated for another 48 hours at 37°C.
3. During the last 5 hours of incubation, plates were labeled with 1 ~Ci 3H
thymidine per well. (New England Nuclear thymidine, catalog # NET-027, 6.7 Ci/mmole).
The _7_ 1 p,Ci was added in 10 pL of PBS (on the day of harvest). The plates were returned to the incubator for the last 5 hours.
4. The radioactive media was removed by aspiration, with care not to disturb the cell monolayer. Then 50p,L of BRL 10X Trypsin was added to each well, followed by incubation at 37°C for 10 minutes or until the monolayer was loosened from the well bottom. Samples were harvested on a glass fiber filter mat using a Skatron 96 well harvester. Mats were counted in a Wallac Betaplate counter.
Results:
Compound l~_50 CCI-779 0.6 ng/mL
Compound III 1.0 ng/mL*
Compound IV 4.0 ng/mL*
* ng/mL equivalent of CCI-779 The results obtained in this standard pharmacological test procedure show that the compounds of this invention inhibit tumor cell growth and are therefore useful as antineoplastic agents. In particular, the compounds of this invention are useful in treating or inhibiting the growth of solid tumors, including sarcomas and carcinomas, such as astrocytomas, prostate cancer, breast cancer, small cell lung cancer, and ovarian cancer.
The compounds of this invention are also useful treatment or inhibition of transplantation rejection such as kidney, heart, liver, lung, bone marrow, pancreas (islet cells), cornea, small bowel, and skin allografts, and heart valve xenografts;
in the treatment or inhibition of graft vs. host disease; in the treatment or inhibition of autoimmune diseases such as lupus, rheumatoid arthritis, diabetes mellitus, myasthenia gravis, and multiple sclerosis; and diseases of inflammation such as psoriasis, dermatitis, eczema, seborrhea, inflammatory bowel disease, pulmonary inflammation (including asthma, chronic obstructive pulmonary disease, emphysema, acute respiratory distress syndrome, bronchitis, and the like) and ocular uveitis;
adult T-cell leukemia/lymphoma; fungal infections; hyperproliferative vascular diseases such as restenosis; graft vascular atherosclerosis; and cardiovascular disease, cerebral vascular disease, and peripheral vascular disease, such as coronary artery disease, cerebrovascular disease, arteriosclerosis, atherosclerosis, nonatheromatous _g_ arteriosclerosis, or vascular wall damage from cellular events leading toward immune mediated vascular damage, and inhibiting stroke or multiinfarct dementia.
When used for restenosis, it is preferred that the compounds of this invention are used to treat restenosis that occurs following an angioplasty procedure. When used for this treating restenosis following an angioplasty, the compounds of this invention can be administered prior to the procedure, during the procedure, subsequent to the procedure, or any combination of the above.
This invention also covers analogous pegylated hydroxyesters of other rapamycins known in the art such as, but not limited to, 29-demethoxyrapamycin, [U.S.
Patent 4,375,464, 32-demethoxyrapamycin under C.A. nomenclature]; rapamycin derivatives in which the double bonds in the 1-, 3-, and/or 5-positions have been reduced [U.S. Patent 5,023,262]; 29-desmethylrapamycin [U.S. Patent 5,093,339, 32-desmethylrapamycin under C.A. nomenclature]; 7,29-bisdesmethylrapamycin [U.S.
Patent 5,093,338, 7,32-desmethylrapamycin under C.A. nomenclature]; 27-hydroxy-rapamycin [U.S. Patent 5,256,790] and 15-hydroxyrapamycin [U.S. Patent 5,102,876].
This invention also covers esters at the 31-position of 42-oxorapamycin [U.S.
Patent 5,023,263]. The disclosures in the above cited U.S. Patents are hereby incorporated by reference.
When administered for the treatment or inhibition of a particular disease state or disorder, it is understood that the effective dosage of the pegylated hydroxyester of rapamycin may vary depending upon the particular compound utilized, the mode of administration, the condition, and severity thereof, of the condition being treated, as well as the various physical factors related to the individual being treated. As used in accordance with invention, satisfactory results may be obtained when the pegylated hydroxyester of rapamycin is administered in a daily oral dosage of from about projected daily dosages of active compound would be 0.1 ~.g/kg - 100 mg/kg, preferably between 0.001 - 25 mg/kg, and more preferably between 0.01 - 5 mg/kg.. The projected daily dosages are expected to vary with route of administration.
Such doses may be administered in any manner useful in directing the active compounds herein to the recipient's bloodstream, including orally, via implants, parenterally (including intravenous, intraperitoneal and subcutaneous injections), _g_ rectally, intranasally, vaginally, and transdermally. The pegylated esters of this invention provide an advantage in ease of formulation and administration over the non-pegylated esters of rapamycin, in that they are significantly more water soluble than corresponding non-pegylated esters. For example, both compounds 3 and 4 had a water solubility of > 100 mg/mL, whereas CCI-779 had a water solubility of < 1.1 ~,g/mL. This advantage is particularly important for oral and parenteral administration.
Oral formulations containing the active compounds of this invention may comprise any conventionally used oral forms, including tablets, capsules, buccal forms, troches, lozenges and oral liquids, suspensions or solutions. Capsules may contain mixtures of the active compounds) with inert fillers and/or diluents such as the pharmaceutically acceptable starches (e.g. corn, potato or tapioca starch), sugars, artificial sweetening agents, powdered celluloses, such as crystalline and microcrystalline celluloses, flours, gelatins, gums, etc. Useful tablet formulations may be made by conventional compression, wet granulation or dry granulation methods and utilize pharmaceutically acceptable diluents, binding agents, lubricants, disintegrants, surface modifying agents (including surfactants), suspending or stabilizing agents, including, but not limited to, magnesium stearate, stearic acid, talc, sodium lauryl sulfate, microcrystalline cellulose, carboxymethylcellulose calcium, polyvinylpyrrolidone, gelatin, alginic acid, acacia gum, xanthan gum, sodium citrate, complex silicates, calcium carbonate, glycine, dextrin, sucrose, sorbitol, dicalcium phosphate, calcium sulfate, lactose, kaolin, mannitol, sodium chloride, talc, dry starches and powdered sugar.
Preferred surface modifying agents include nonionic and anionic surface modifying agents. Representative examples of surface modifying agents include, but are not limited to, poloxamer 188, benzalkonium chloride, calcium stearate, cetostearl alcohol, cetomacrogol emulsifying wax, sorbitan esters, colloidol silicon dioxide, phosphates, sodium dodecylsulfate, magnesium aluminum silicate, and triethanolamine. Oral formulations herein may utilize standard delay or time release formulations to alter the absorption of the active compound(s). The oral formulation may also consist of administering the active ingredient in water or a fruit juice, containing appropriate solubilizers or emulsifiers as needed.
In some cases it may be desirable to administer the compounds directly to the airways in the form of an aerosol.
The compounds of this invention may also be administered parenterally or intraperitoneally. Solutions or suspensions of these active compounds as a free base or pharmacologically acceptable salt can be prepared in water suitably mixed with a surfactant such as hydroxy-propylcellulose. Dispersions can also be prepared in glycerol, liquid polyethylene glycols and mixtures thereof in oils. Under ordinary conditions of storage and use, these preparation contain a preservative to prevent the growth of microorganisms.
The pharmaceutical forms suitable for injectable use include sterile aqueous solutions or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersions. In all cases, the form must be sterile and must be fluid to the extent that easy syringability exists. It must be stable under the conditions of manufacture and storage and must be preserved against the contaminating action of microorganisms such as bacteria and fungi. The carrier can be a solvent or dispersion medium containing, for example, water, ethanol, polyol (e.g., glycerol, propylene glycol and liquid polyethylene glycol), suitable mixtures thereof, and vegetable oils.
For the purposes of this disclosure, transdermal administrations are understood to include all administrations across the surface of the body and the inner linings of bodily passages including epithelial and mucosal tissues. Such administrations may be carried out using the present compounds, or pharmaceutically acceptable salts thereof, in lotions, creams, foams, patches, suspensions, solutions, and suppositories (rectal and vaginal).
Transdermal administration may be accomplished through the use of a transdermal patch containing the active compound and a carrier that is inert to the active compound, is non toxic to the skin, and allows delivery of the agent for systemic absorption into the blood stream via the skin. The carrier may take any number of forms such as creams and ointments, pastes, gels, and occlusive devices. The creams and ointments may be viscous liquid or semisolid emulsions of either the oil-in-water or water-in-oil type. Pastes comprised of absorptive powders dispersed in petroleum or hydrophilic petroleum containing the active ingredient may also be suitable. A
variety of occlusive devices may be used to release the active ingredient into the blood stream such as a semi-permeable membrane covering a reservoir containing the active ingredient with or without a carrier, or a matrix containing the active ingredient. Other occlusive devices are known in the literature.
Suppository formulations may be made from traditional materials, including cocoa butter, with or without the addition of waxes to alter the suppository's melting point, and glycerin. Water soluble suppository bases, such as polyethylene glycols of various molecular weights, may also be used.
The preparation of representative examples of this invention is described below.
Example 1 Preparation of Rapamycin 42-ester with 3-hydroxy-2-(2-iodo-acetoxymethyl)-2-methyl propionic acid (Compound I) and Rapamycin 42-ester with 3-(2-lodo-acetoxy)-2-(2-iodo acetoxymethyl)-2-methyl-propionic acid (Compound II) O~I O~I
O O
O O
.... ~ ~O
Compound I Compound II
CCI-779 (1.03 g 1.0 x 10-3 mole), 4-Dimethylaminopyridine (3.0 mg) and 1,3-Dicyclohexylcarbodiimide (0.136 g, 6.6 x 10-4 mole) were dissolved in 50 mL
anhydrous methylene chloride in a 250 mL round-bottom flask. lodoacetic acid (0.185 g, 1.0 x 10-3 mole) was dissolved in 10 mL anhydrous methylene chloride. The iodoacetic' acid solution was added into reaction mixture over a period of 10 min with stirring by a magnetic bar. Then the reaction mixture was stirred at room temperature for another 2.5 h. The solution was then filtered through a filter paper. The filtrate was transferred to a separatory funnel, washed 50 mL of sodium bicarbonate solution (5.5 g/ 100 mL) and then washed with 2 x 50 mL of water. The methylene chloride layer was dried with 10 g anhydrous sodium sulfate for 2 h. Then sodium sulfate was filtered out and methylene chloride was removed by rotary evaporation. A total of 0.93 g yellow solid was obtained.
Isolation of pure compound I and compound II was performed by preparative HPLC
on a Prep Nova-pak HR C18 (300 x 19 mm) column from Waters. Compound I eluted at 18.4 min and compound II eluted at 24.4 min using a gradient ( 30% A, 70% B for 5 min. then to 100% B in 30 min.). A is 90% water, 10% acetonitrile; B is 10% water, 90%
acetonitrile. The fraction was collected and extracted by 2 x 100 ml methylene chloride.
The organic layer was combined and dried with anhydrous sodium sulfate for 4 h. The organic solvent was removed by rotary evaporation to dryness. Compound I, a yellowish solid was obtained (0.14 g). 'H NMR (CDCL3, 400 MHz) S 3.68 (s, 2H, I-CH2-C02-), 4.28 (dd, 2H, -C02-CH2-). MS m/z 1215.8 (M+NH4)+. Compound II, a yellowish solid was obtained (0.08 g).'H NMR (CDCL3, 400 MHz) 8 3.72 (s, 4H, 2 x I-CH2-C02-), 4.28 (dd, 4H, 2 x -C02-CH -). MS m/z 1383.6 (M+NH4)+.
Example 2 Preparation of CCI-779-PEG 5000 -Rapamycin 42-ester with 3-hydroxy-2-(hydroxymethyl)-2-methylpropionic acid methoxypoly(ethylene glycol) thiol 5000 conjugate (Compound III) ~S ~O~OCH3 n (Compound III) with average n = 108 Compound I (90 mg, 7.6 x 10-5 mole) was dissolved in 40 mL of solution containing 50% acetonitrile and 50% aqueous NaHC03 (0.1 M) solution. The solution was flushed with N2 for 10 min. The original sample 10 p,L was taken for HPLC
analysis.
Then mPEG-SH 5000 (450 mg, 9.1 x 10-5 mole) was added to the reaction solution and the reaction mixture was stirred at room temperature for another 45 min. The reaction was checked again by taking 10 p,L sample for HPLC analysis. The chromatogram showed that compound I was 100% converted to Compound III. The reaction mixture was extracted with 2 x 100 mL methylene chloride. The organic layer was dried with anhydrous sodium sulfate then filtered. The filtrate was concentrated to a volume of 20 mL by rotaty evaporation. The crude product was precipitated out after adding 150 mL
ether. A total of 404 mg white powder was obtained after filtered out by a sintered glass funnel and dried under vacuum. Isolation of pure compound 111, which may also be referred to as CCI-779-PEG 5000, was performed by preparative HPLC on a Prep Nova-pak HR C18 (300 x 19 mm) column from Waters. Compound III eluted at 18 min using a gradient (60% A, 40% B for 5 min then at 20% A, 80% B in 30 min). The fraction was collected and extracted by 2 x 100 mL methylene chloride. The organic layer was combined and dried with anhydrous sodium sulfate for 4 hr. The organic solvent was removed by rotary evaporation to dryness. The residue was dissolved in 5 mL
methylene chloride and was precipitated out after adding 150 mL ether. A white powder was obtained after filtered out by a sintered glass funnel and dried under vacuum. 'H
NMR (CDCI3, 400 MHz) ~ 2.83 (t, 2H, -S-CH2-CH2-), 3.30 (s, 2H, -CO-CH2-S), 3.38 (s, 3H, -OCH3), 4.25 (dd, 2H, -C02-CH2-). MS (MALD/TOF) m/z 5894.5 (ave. M. Wt.) which indicate the average n=108 for compound III, with 99% of n being between 65 and 155.
Example 3 Preparation of CCI-779-(PEG 5000)2 -Rapamycin 42-ester with 3-hydroxy-2-(hydroxymethyl)-2-methylpropionic acid bis(methoxypoly(ethylene glycol) thiol 5000) conjugate (Compound IV) s o~/~
O ~ - / nOCH3 O
O
~O O
~mOCH3 ~S~O~'nOCH3 ."...
~,,o N~O O I OH
IOI ___ _m~ ,O
CCI-779 -(PEG 5000)2 (Compound IV) with average n = 108 Compound II (50 mg, 3.6 x 10-5 mole) was dissolved in 30 mL of solution containing 50% acetonitrile and 50% aqueous NaHC03 (0.1 M) solution. The solution was flushed with N2 for 10 min. The original sample 10 ~L was taken for HPLC
analysis.
Then mPEG-SH 5000 (500 mg, 1.0 x 10-4 mole) was added to the reaction solution and the reaction mixture was stirred at room temperature for about 70 min. The reaction was checked again by taking 10 g,L sample for HPLC analysis. The chromatogram showed that compound II was 100% converted to Compound IV. The reaction mixture was extracted with 2 x 50 mL methylene chloride. The organic layer was dried with anhydrous sodium sulfate then filtered. The filtrate was concentrated to a volume of 10 mL by rotaty evaporation. The crude product was precipitated out after adding 150 mL
ether. A total of 500 mg white powder was obtained after filtered out by a sintered glass funnel and dried under vacuum. Isolation of pure compound IV, which may also be referred to as CCI-779-(PEG 5000)2, was performed by preparative HPLC on a Prep Nova-pak HR C18 (300 x 19 mm) column from Waters. Compound IV eluted at 22 min using a gradient (60% A, 40% B for 5 min then at 20% A, 80% B in 30 min). The fraction was collected and extracted by 2 x 100 mL methylene chloride. The organic layer was combined and dried with anhydrous sodium sulfate for 4 hr. The organic solvent was removed by rotary evaporation to dryness. The residue was dissolved in 5 mL
methylene chloride and was precipitated out after adding 150 mL ether. A white powder was obtained after filtered out by a sintered glass funnel and dried under vacuum. 'H
NMR (CDCI3, 400 MHz) 8 2.81 (t, 4H, 2 x S-CH2-CH2-), 3.29 (s, 4H, 2 x CO-CH2-S-), 3.38 (s, 6H, 2 x -OCH3), 4.26 (dd, 4H, 2 x -C02-CH2-). MS (MALD/TOF) m/z 10760.7 (ave. M. Wt.) which indicate the average n=108 for compound IV, with 99% of n being between 65 and 155.
The terms "alkyl", "alkenyl" and "alkynyl" include both straight chain as well as branched carbon chains. Alkyl groups may contain 1-6, e.g. 1-4 carbon atoms, eg methyl, ethyl, propyl and tert-butyl. Alkenyl or alkynyl may contain 2-7 carbon atoms eg 2- 5 such as vinyl, propenyl or acetylenyl.
This invention also provides a process for preparing compounds of formula (A) which comprises reacting a compound of formula (B):
ORA
~'OMe ., O O ~ ORB
3i O O O Me0'' O
HO
'O OMe '~ ~ / ~ /
I ' wherein RA is R1 as hereinbefore defined, and wherein RB is R2 as hereinbefore defined, and in which R10 is as hereinbefore defined in connection with formula (A) providing that at least one R10 group represents -COCH2-X where X is a suitable leaving group;
with a compound of formula (C):
HS-CH2CH2-(O-CH2-CH2)n-OCH3 (C) wherein n is as defined above, if necessary any reactive substituent group being protected and removing any protecting group as required, to give a compound of formula (A) or a pharmaceutically acceptable salt thereof.
Compounds of formula (B) also form another aspect of this invention.
The reagents used in the preparation of the compounds of this invention can be either commercially obtained or can be prepared by standard procedures described in the literature.
The preparation of the hydroxyesters of rapamycin, from which the pegylated hydroxyesters are made from, are described in US Patent 5,362,718 and WO
95/28406, which is hereby incorporated by reference. Conveniently one or more of the hydroxyesters may be acylated with a acylating agent having the general structure X-CH2CO~H, where X is a suitable leaving group, such as bromine or iodine, in the presence of a coupling agent such as dicyclohexylcarbodiimide (DCC) in the presence of a base catalyst, such as dimethylaminopyridine (DMAP). Accordingly, rapamycin ester with 3-hydroxy-2-(2-iodo-acetoxymethyl)-2-methyl-propionic acid (Compound I) and rapamycin 42-ester with 3-(2-lodo-acetoxy)-2-(2-iodo-acetoxymethyl)-2-methyl propionic acid (Compound II) are intermediates prepared using this methodology, that are useful in the production of the corresponding pegylated hydroxyesters of rapamycin (Compounds III and IV, respectively).
The pegylated esters may be conveniently be produced utilizing the polyethylene glycols known in the art, such as those described on pages 355 to 361 of the Handbook of Pharmaceutical Excipients, Second Edition, 1994. The preferred compounds of this invention may also be described as those of the formula esterified using polyethylene glycols having an average molecular weight of from about 200 to about 200,000. A preferred range of the PEG esters of this invention includes those in which the molecular weight of the polyethylene glycol portion of the ester chain has a molecular weight in the range of from about 300 to about 20,000, more preferably between about 350 and about 6,000. Reaction of the hydroxyester of rapamycin which has been acylated as described above, with a suitable polyethyleneglycol thiol in the presence of a base, such as sodium bicarbonate provides the desired pegylated hydroxyester of rapamycin.
The antitumor activity of the compounds of this invention were confirmed in a standard pharmacological test procedure which measures the inhibition of U87MG
human glioblastoma cell growth (as a function of 3H-thymidine incorporation), using CCI-779-PEG 5000 ester (Compound III) and CCI-779-(PEG 5000)2 ester (Compound IV) as representative compounds of this invention. The following briefly describes the procedure used and results obtained. U87MG human glioblastoma cells (ATCC #
HTB-14; available from the American Type Culture Collection; 10801 University Boulevard;
Manassas, VA 20110;), were grown in the following media.
Growth Medium: BRL Minimum Essential Medium with Earle Salts (500 mL) + 5 mL BRL MEM Non-Essential Amino Acids (lOmM) +5 mL BRL Penicillin-Streptomycin ( 10000 u/m L,10000 ~g/m L) +5 mL BRL Na Pyruvate Solution (100mM) +5 mL BRL L-Glutamine 200mM
+50 mL BRL Fetal Bovine Serum (Qualified) Test Procedure:
1. Cells were trypsinized and plated at a concentration of 104 cells/well in a final volume of 200 pL growth medium in 96-well flat bottom plates and allowed to adhere for 24 hours at 37°C.
2. The media was removed by aspiration with care to not disturb the cell monolayer.
200 p,L of fresh growth media was added per well, allowing enough wells for samples to be run in triplicate. Test compounds were added in 10 p.L phosphate buffer solution (PBS) and incubated for another 48 hours at 37°C.
3. During the last 5 hours of incubation, plates were labeled with 1 ~Ci 3H
thymidine per well. (New England Nuclear thymidine, catalog # NET-027, 6.7 Ci/mmole).
The _7_ 1 p,Ci was added in 10 pL of PBS (on the day of harvest). The plates were returned to the incubator for the last 5 hours.
4. The radioactive media was removed by aspiration, with care not to disturb the cell monolayer. Then 50p,L of BRL 10X Trypsin was added to each well, followed by incubation at 37°C for 10 minutes or until the monolayer was loosened from the well bottom. Samples were harvested on a glass fiber filter mat using a Skatron 96 well harvester. Mats were counted in a Wallac Betaplate counter.
Results:
Compound l~_50 CCI-779 0.6 ng/mL
Compound III 1.0 ng/mL*
Compound IV 4.0 ng/mL*
* ng/mL equivalent of CCI-779 The results obtained in this standard pharmacological test procedure show that the compounds of this invention inhibit tumor cell growth and are therefore useful as antineoplastic agents. In particular, the compounds of this invention are useful in treating or inhibiting the growth of solid tumors, including sarcomas and carcinomas, such as astrocytomas, prostate cancer, breast cancer, small cell lung cancer, and ovarian cancer.
The compounds of this invention are also useful treatment or inhibition of transplantation rejection such as kidney, heart, liver, lung, bone marrow, pancreas (islet cells), cornea, small bowel, and skin allografts, and heart valve xenografts;
in the treatment or inhibition of graft vs. host disease; in the treatment or inhibition of autoimmune diseases such as lupus, rheumatoid arthritis, diabetes mellitus, myasthenia gravis, and multiple sclerosis; and diseases of inflammation such as psoriasis, dermatitis, eczema, seborrhea, inflammatory bowel disease, pulmonary inflammation (including asthma, chronic obstructive pulmonary disease, emphysema, acute respiratory distress syndrome, bronchitis, and the like) and ocular uveitis;
adult T-cell leukemia/lymphoma; fungal infections; hyperproliferative vascular diseases such as restenosis; graft vascular atherosclerosis; and cardiovascular disease, cerebral vascular disease, and peripheral vascular disease, such as coronary artery disease, cerebrovascular disease, arteriosclerosis, atherosclerosis, nonatheromatous _g_ arteriosclerosis, or vascular wall damage from cellular events leading toward immune mediated vascular damage, and inhibiting stroke or multiinfarct dementia.
When used for restenosis, it is preferred that the compounds of this invention are used to treat restenosis that occurs following an angioplasty procedure. When used for this treating restenosis following an angioplasty, the compounds of this invention can be administered prior to the procedure, during the procedure, subsequent to the procedure, or any combination of the above.
This invention also covers analogous pegylated hydroxyesters of other rapamycins known in the art such as, but not limited to, 29-demethoxyrapamycin, [U.S.
Patent 4,375,464, 32-demethoxyrapamycin under C.A. nomenclature]; rapamycin derivatives in which the double bonds in the 1-, 3-, and/or 5-positions have been reduced [U.S. Patent 5,023,262]; 29-desmethylrapamycin [U.S. Patent 5,093,339, 32-desmethylrapamycin under C.A. nomenclature]; 7,29-bisdesmethylrapamycin [U.S.
Patent 5,093,338, 7,32-desmethylrapamycin under C.A. nomenclature]; 27-hydroxy-rapamycin [U.S. Patent 5,256,790] and 15-hydroxyrapamycin [U.S. Patent 5,102,876].
This invention also covers esters at the 31-position of 42-oxorapamycin [U.S.
Patent 5,023,263]. The disclosures in the above cited U.S. Patents are hereby incorporated by reference.
When administered for the treatment or inhibition of a particular disease state or disorder, it is understood that the effective dosage of the pegylated hydroxyester of rapamycin may vary depending upon the particular compound utilized, the mode of administration, the condition, and severity thereof, of the condition being treated, as well as the various physical factors related to the individual being treated. As used in accordance with invention, satisfactory results may be obtained when the pegylated hydroxyester of rapamycin is administered in a daily oral dosage of from about projected daily dosages of active compound would be 0.1 ~.g/kg - 100 mg/kg, preferably between 0.001 - 25 mg/kg, and more preferably between 0.01 - 5 mg/kg.. The projected daily dosages are expected to vary with route of administration.
Such doses may be administered in any manner useful in directing the active compounds herein to the recipient's bloodstream, including orally, via implants, parenterally (including intravenous, intraperitoneal and subcutaneous injections), _g_ rectally, intranasally, vaginally, and transdermally. The pegylated esters of this invention provide an advantage in ease of formulation and administration over the non-pegylated esters of rapamycin, in that they are significantly more water soluble than corresponding non-pegylated esters. For example, both compounds 3 and 4 had a water solubility of > 100 mg/mL, whereas CCI-779 had a water solubility of < 1.1 ~,g/mL. This advantage is particularly important for oral and parenteral administration.
Oral formulations containing the active compounds of this invention may comprise any conventionally used oral forms, including tablets, capsules, buccal forms, troches, lozenges and oral liquids, suspensions or solutions. Capsules may contain mixtures of the active compounds) with inert fillers and/or diluents such as the pharmaceutically acceptable starches (e.g. corn, potato or tapioca starch), sugars, artificial sweetening agents, powdered celluloses, such as crystalline and microcrystalline celluloses, flours, gelatins, gums, etc. Useful tablet formulations may be made by conventional compression, wet granulation or dry granulation methods and utilize pharmaceutically acceptable diluents, binding agents, lubricants, disintegrants, surface modifying agents (including surfactants), suspending or stabilizing agents, including, but not limited to, magnesium stearate, stearic acid, talc, sodium lauryl sulfate, microcrystalline cellulose, carboxymethylcellulose calcium, polyvinylpyrrolidone, gelatin, alginic acid, acacia gum, xanthan gum, sodium citrate, complex silicates, calcium carbonate, glycine, dextrin, sucrose, sorbitol, dicalcium phosphate, calcium sulfate, lactose, kaolin, mannitol, sodium chloride, talc, dry starches and powdered sugar.
Preferred surface modifying agents include nonionic and anionic surface modifying agents. Representative examples of surface modifying agents include, but are not limited to, poloxamer 188, benzalkonium chloride, calcium stearate, cetostearl alcohol, cetomacrogol emulsifying wax, sorbitan esters, colloidol silicon dioxide, phosphates, sodium dodecylsulfate, magnesium aluminum silicate, and triethanolamine. Oral formulations herein may utilize standard delay or time release formulations to alter the absorption of the active compound(s). The oral formulation may also consist of administering the active ingredient in water or a fruit juice, containing appropriate solubilizers or emulsifiers as needed.
In some cases it may be desirable to administer the compounds directly to the airways in the form of an aerosol.
The compounds of this invention may also be administered parenterally or intraperitoneally. Solutions or suspensions of these active compounds as a free base or pharmacologically acceptable salt can be prepared in water suitably mixed with a surfactant such as hydroxy-propylcellulose. Dispersions can also be prepared in glycerol, liquid polyethylene glycols and mixtures thereof in oils. Under ordinary conditions of storage and use, these preparation contain a preservative to prevent the growth of microorganisms.
The pharmaceutical forms suitable for injectable use include sterile aqueous solutions or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersions. In all cases, the form must be sterile and must be fluid to the extent that easy syringability exists. It must be stable under the conditions of manufacture and storage and must be preserved against the contaminating action of microorganisms such as bacteria and fungi. The carrier can be a solvent or dispersion medium containing, for example, water, ethanol, polyol (e.g., glycerol, propylene glycol and liquid polyethylene glycol), suitable mixtures thereof, and vegetable oils.
For the purposes of this disclosure, transdermal administrations are understood to include all administrations across the surface of the body and the inner linings of bodily passages including epithelial and mucosal tissues. Such administrations may be carried out using the present compounds, or pharmaceutically acceptable salts thereof, in lotions, creams, foams, patches, suspensions, solutions, and suppositories (rectal and vaginal).
Transdermal administration may be accomplished through the use of a transdermal patch containing the active compound and a carrier that is inert to the active compound, is non toxic to the skin, and allows delivery of the agent for systemic absorption into the blood stream via the skin. The carrier may take any number of forms such as creams and ointments, pastes, gels, and occlusive devices. The creams and ointments may be viscous liquid or semisolid emulsions of either the oil-in-water or water-in-oil type. Pastes comprised of absorptive powders dispersed in petroleum or hydrophilic petroleum containing the active ingredient may also be suitable. A
variety of occlusive devices may be used to release the active ingredient into the blood stream such as a semi-permeable membrane covering a reservoir containing the active ingredient with or without a carrier, or a matrix containing the active ingredient. Other occlusive devices are known in the literature.
Suppository formulations may be made from traditional materials, including cocoa butter, with or without the addition of waxes to alter the suppository's melting point, and glycerin. Water soluble suppository bases, such as polyethylene glycols of various molecular weights, may also be used.
The preparation of representative examples of this invention is described below.
Example 1 Preparation of Rapamycin 42-ester with 3-hydroxy-2-(2-iodo-acetoxymethyl)-2-methyl propionic acid (Compound I) and Rapamycin 42-ester with 3-(2-lodo-acetoxy)-2-(2-iodo acetoxymethyl)-2-methyl-propionic acid (Compound II) O~I O~I
O O
O O
.... ~ ~O
Compound I Compound II
CCI-779 (1.03 g 1.0 x 10-3 mole), 4-Dimethylaminopyridine (3.0 mg) and 1,3-Dicyclohexylcarbodiimide (0.136 g, 6.6 x 10-4 mole) were dissolved in 50 mL
anhydrous methylene chloride in a 250 mL round-bottom flask. lodoacetic acid (0.185 g, 1.0 x 10-3 mole) was dissolved in 10 mL anhydrous methylene chloride. The iodoacetic' acid solution was added into reaction mixture over a period of 10 min with stirring by a magnetic bar. Then the reaction mixture was stirred at room temperature for another 2.5 h. The solution was then filtered through a filter paper. The filtrate was transferred to a separatory funnel, washed 50 mL of sodium bicarbonate solution (5.5 g/ 100 mL) and then washed with 2 x 50 mL of water. The methylene chloride layer was dried with 10 g anhydrous sodium sulfate for 2 h. Then sodium sulfate was filtered out and methylene chloride was removed by rotary evaporation. A total of 0.93 g yellow solid was obtained.
Isolation of pure compound I and compound II was performed by preparative HPLC
on a Prep Nova-pak HR C18 (300 x 19 mm) column from Waters. Compound I eluted at 18.4 min and compound II eluted at 24.4 min using a gradient ( 30% A, 70% B for 5 min. then to 100% B in 30 min.). A is 90% water, 10% acetonitrile; B is 10% water, 90%
acetonitrile. The fraction was collected and extracted by 2 x 100 ml methylene chloride.
The organic layer was combined and dried with anhydrous sodium sulfate for 4 h. The organic solvent was removed by rotary evaporation to dryness. Compound I, a yellowish solid was obtained (0.14 g). 'H NMR (CDCL3, 400 MHz) S 3.68 (s, 2H, I-CH2-C02-), 4.28 (dd, 2H, -C02-CH2-). MS m/z 1215.8 (M+NH4)+. Compound II, a yellowish solid was obtained (0.08 g).'H NMR (CDCL3, 400 MHz) 8 3.72 (s, 4H, 2 x I-CH2-C02-), 4.28 (dd, 4H, 2 x -C02-CH -). MS m/z 1383.6 (M+NH4)+.
Example 2 Preparation of CCI-779-PEG 5000 -Rapamycin 42-ester with 3-hydroxy-2-(hydroxymethyl)-2-methylpropionic acid methoxypoly(ethylene glycol) thiol 5000 conjugate (Compound III) ~S ~O~OCH3 n (Compound III) with average n = 108 Compound I (90 mg, 7.6 x 10-5 mole) was dissolved in 40 mL of solution containing 50% acetonitrile and 50% aqueous NaHC03 (0.1 M) solution. The solution was flushed with N2 for 10 min. The original sample 10 p,L was taken for HPLC
analysis.
Then mPEG-SH 5000 (450 mg, 9.1 x 10-5 mole) was added to the reaction solution and the reaction mixture was stirred at room temperature for another 45 min. The reaction was checked again by taking 10 p,L sample for HPLC analysis. The chromatogram showed that compound I was 100% converted to Compound III. The reaction mixture was extracted with 2 x 100 mL methylene chloride. The organic layer was dried with anhydrous sodium sulfate then filtered. The filtrate was concentrated to a volume of 20 mL by rotaty evaporation. The crude product was precipitated out after adding 150 mL
ether. A total of 404 mg white powder was obtained after filtered out by a sintered glass funnel and dried under vacuum. Isolation of pure compound 111, which may also be referred to as CCI-779-PEG 5000, was performed by preparative HPLC on a Prep Nova-pak HR C18 (300 x 19 mm) column from Waters. Compound III eluted at 18 min using a gradient (60% A, 40% B for 5 min then at 20% A, 80% B in 30 min). The fraction was collected and extracted by 2 x 100 mL methylene chloride. The organic layer was combined and dried with anhydrous sodium sulfate for 4 hr. The organic solvent was removed by rotary evaporation to dryness. The residue was dissolved in 5 mL
methylene chloride and was precipitated out after adding 150 mL ether. A white powder was obtained after filtered out by a sintered glass funnel and dried under vacuum. 'H
NMR (CDCI3, 400 MHz) ~ 2.83 (t, 2H, -S-CH2-CH2-), 3.30 (s, 2H, -CO-CH2-S), 3.38 (s, 3H, -OCH3), 4.25 (dd, 2H, -C02-CH2-). MS (MALD/TOF) m/z 5894.5 (ave. M. Wt.) which indicate the average n=108 for compound III, with 99% of n being between 65 and 155.
Example 3 Preparation of CCI-779-(PEG 5000)2 -Rapamycin 42-ester with 3-hydroxy-2-(hydroxymethyl)-2-methylpropionic acid bis(methoxypoly(ethylene glycol) thiol 5000) conjugate (Compound IV) s o~/~
O ~ - / nOCH3 O
O
~O O
~mOCH3 ~S~O~'nOCH3 ."...
~,,o N~O O I OH
IOI ___ _m~ ,O
CCI-779 -(PEG 5000)2 (Compound IV) with average n = 108 Compound II (50 mg, 3.6 x 10-5 mole) was dissolved in 30 mL of solution containing 50% acetonitrile and 50% aqueous NaHC03 (0.1 M) solution. The solution was flushed with N2 for 10 min. The original sample 10 ~L was taken for HPLC
analysis.
Then mPEG-SH 5000 (500 mg, 1.0 x 10-4 mole) was added to the reaction solution and the reaction mixture was stirred at room temperature for about 70 min. The reaction was checked again by taking 10 g,L sample for HPLC analysis. The chromatogram showed that compound II was 100% converted to Compound IV. The reaction mixture was extracted with 2 x 50 mL methylene chloride. The organic layer was dried with anhydrous sodium sulfate then filtered. The filtrate was concentrated to a volume of 10 mL by rotaty evaporation. The crude product was precipitated out after adding 150 mL
ether. A total of 500 mg white powder was obtained after filtered out by a sintered glass funnel and dried under vacuum. Isolation of pure compound IV, which may also be referred to as CCI-779-(PEG 5000)2, was performed by preparative HPLC on a Prep Nova-pak HR C18 (300 x 19 mm) column from Waters. Compound IV eluted at 22 min using a gradient (60% A, 40% B for 5 min then at 20% A, 80% B in 30 min). The fraction was collected and extracted by 2 x 100 mL methylene chloride. The organic layer was combined and dried with anhydrous sodium sulfate for 4 hr. The organic solvent was removed by rotary evaporation to dryness. The residue was dissolved in 5 mL
methylene chloride and was precipitated out after adding 150 mL ether. A white powder was obtained after filtered out by a sintered glass funnel and dried under vacuum. 'H
NMR (CDCI3, 400 MHz) 8 2.81 (t, 4H, 2 x S-CH2-CH2-), 3.29 (s, 4H, 2 x CO-CH2-S-), 3.38 (s, 6H, 2 x -OCH3), 4.26 (dd, 4H, 2 x -C02-CH2-). MS (MALD/TOF) m/z 10760.7 (ave. M. Wt.) which indicate the average n=108 for compound IV, with 99% of n being between 65 and 155.
Claims (32)
1. A compound having the structure (A) wherein R1 and R2 are each, independently, hydrogen or -CO(CR3R4)b(CR5R6)dCR7R8R9;
R3 and R4 are each, independently, hydrogen, alkyl of 1-6 carbon atoms, alkenyl of 2-7 carbon atoms, alkynyl of 2-7 carbon atoms, trifluoromethyl, or -F;
R5 and R6 are each, independently, hydrogen, alkyl of 1-6 carbon atoms, alkenyl of 2-7 carbon atoms, alkynyl of 2-7 carbon atoms, -(CR3R4)fOR10, -CF3, -F, or -CO2R11;
R7 is hydrogen, alkyl of 1-6 carbon atoms, alkenyl of 2-7 carbon atoms, alkynyl of 2-7 carbon atoms, -(CR3R4)fOR10, -CF3, -F, or -CO2R11;
R8 and R9 are each, independently, hydrogen, alkyl of 1-6 carbon atoms, alkenyl of 2-7 carbon atoms, alkynyl of 2-7 carbon atoms, -(CR3R4)fOR10, -CF3, -F, or -CO2R11;
R10 is hydrogen or -COCH2-S-CH2CH2-(O-CH2-CH2)n-OCH3;
R11 is hydrogen, alkyl of 1-6 carbon atoms, alkenyl of 2-7 carbon atoms, alkynyl of 2-7 carbon atoms, or phenylalkyl of 7-10 carbon atoms;
b = 0-6;
d = 0-6;
f = 0-6;
n = 5-450;
with the proviso that R1 and R2 are both not hydrogen and further provided that either R1 or R2 contains at least one -(CR3R4)fOR10 group in which R10 is -COCH2-S-CH2CH2-(O-CH2-CH2)n-OCH3, or a pharmaceutically acceptable salt thereof.
R3 and R4 are each, independently, hydrogen, alkyl of 1-6 carbon atoms, alkenyl of 2-7 carbon atoms, alkynyl of 2-7 carbon atoms, trifluoromethyl, or -F;
R5 and R6 are each, independently, hydrogen, alkyl of 1-6 carbon atoms, alkenyl of 2-7 carbon atoms, alkynyl of 2-7 carbon atoms, -(CR3R4)fOR10, -CF3, -F, or -CO2R11;
R7 is hydrogen, alkyl of 1-6 carbon atoms, alkenyl of 2-7 carbon atoms, alkynyl of 2-7 carbon atoms, -(CR3R4)fOR10, -CF3, -F, or -CO2R11;
R8 and R9 are each, independently, hydrogen, alkyl of 1-6 carbon atoms, alkenyl of 2-7 carbon atoms, alkynyl of 2-7 carbon atoms, -(CR3R4)fOR10, -CF3, -F, or -CO2R11;
R10 is hydrogen or -COCH2-S-CH2CH2-(O-CH2-CH2)n-OCH3;
R11 is hydrogen, alkyl of 1-6 carbon atoms, alkenyl of 2-7 carbon atoms, alkynyl of 2-7 carbon atoms, or phenylalkyl of 7-10 carbon atoms;
b = 0-6;
d = 0-6;
f = 0-6;
n = 5-450;
with the proviso that R1 and R2 are both not hydrogen and further provided that either R1 or R2 contains at least one -(CR3R4)fOR10 group in which R10 is -COCH2-S-CH2CH2-(O-CH2-CH2)n-OCH3, or a pharmaceutically acceptable salt thereof.
2. A compound according to claim 1 wherein R2 is hydrogen.
3. A compound according to claim 1 or claim 2 wherein b=0 and d=0.
4. A compound according to any one of claims 1 to 3 wherein f=1.
5. A compound according to any one of claims 1 to 4 wherein R3 and R4 are hydrogen.
6. A compound according to any one of claims 1 to 5 wherein R7 is hydrogen, alkyl of 1-6 carbon atoms, alkenyl of 2-7 carbon atoms, alkynyl of 2-7 carbon atoms or -CF3.
7. A compound according to any one of claims 1 to 6 wherein R8 and R9 are -(CR3R4)fOR10.
8. A compound according to any one of claims 1 to 7 wherein n = 5 - 200.
9. A compound according to claim 8 wherein n = 8 - 135.
10. A compound according to claim 8 wherein n = 8 - 20.
11. A compound according to claim 8 wherein n = 90 - 120.
12. A compound according to claim 1, which is rapamycin 42-ester with 3-hydroxy-2-(hydroxymethyl)-2-methylpropionic acid methoxypoly(ethylene glycol) thiol 5000 conjugate.
13. A compound according to claim 1, which is rapamycin 42-ester with 3-hydroxy-2-(hydroxymethyl)-2-methylpropionic acid bis(methoxypoly(ethylene glycol) thiol 5000) conjugate.
14. A method of treating or inhibiting transplant rejection or graft vs. host disease in a mammal in need thereof, which comprises providing to said mammal an effective amount of a compound of formula (A) as claimed in any one of claims 1 to 13.
15. A method of treating or inhibiting a solid tumor in a mammal in need thereof, which comprises providing to said mammal a compound of formula (A) as claimed in any one of claims 1 to 13.
16. A method of treating or inhibiting a fungal infection in a mammal in need thereof, which comprises providing to said mammal an effective amount of a compound of formula (A) as claimed in any one of claims 1 to 13.
17. A method of treating or inhibiting rheumatoid arthritis in a mammal in need thereof, which comprises providing to said mammal an effective amount of a compound of formula (A) as claimed in any one of claims 1 to 13.
18. A method of treating or inhibiting multiple sclerosis in a mammal in need thereof, which comprises providing to said mammal an effective amount of a compound of formula (A) as claimed in any one of claims 1 to 13.
19. A method of treating or inhibiting restenosis in a mammal in need thereof, which comprises providing to said mammal an effective amount of a compound of formula (A) as claimed in any one of claims 1 to 13.
20. A method of treating or inhibiting pulmonary inflammation in a mammal in need thereof, which comprises providing to said mammal an effective amount of a compound of formula (A) as claimed in any one of claims 1 to 13.
21. Use of a compound of formula (A) as claimed in any one of claims 1 to 13 or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating or inhibiting transplant rejection or graft vs. host disease in a mammal.
22. Use of a compound of formula (A) as claimed in any one of claims 1 to 13 or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating or inhibiting a solid tumor in a mammal.
23. Use of a compound of formula (A) as claimed in any one of claims 1 to 13 or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating or inhibiting a fungal infection in a mammal.
24. Use of a compound of formula (A) as claimed in any one of claims 1 to 13 or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating or inhibiting rheumatoid arthritis in a mammal.
25. Use of a compound of formula (A) as claimed in any one of claims 1 to 13 or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating or inhibiting multiple sclerosis in a mammal.
26. Use of a compound of formula (A) as claimed in any one of claims 1 to 13 or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating or inhibiting restenosis in a mammal.
27. Use of a compound of formula (A) as claimed in any one of claims 1 to 13 or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating or inhibiting pulmonary inflammation in a mammal.
28. A pharmaceutical composition which comprises a compound of the formula (A) as claimed in anyone of claims 1 to 13 and a pharmaceutically acceptable carrier.
29. A process for the preparation of a compound of the formula (A) as defined in claim 1 which comprises reacting a compound of formula (B) wherein R A is R1 as defined in claim 1, and wherein R B is R2 as defined in claim 1, and in which R10 is as defined in connection with formula (A) in claim 1, providing that at least one R10 group represents -COCH2-X where X is a suitable leaving group;
with a compound of formula (C):
HS-CH2CH2-(O-CH2-CH2)n-OCH3 (C) wherein n is as defined in claim 1, if necessary any reactive substituent group being protected and removing any protecting group as required to give a corresponding compound of formula (A).
with a compound of formula (C):
HS-CH2CH2-(O-CH2-CH2)n-OCH3 (C) wherein n is as defined in claim 1, if necessary any reactive substituent group being protected and removing any protecting group as required to give a corresponding compound of formula (A).
30. A compound of formula (B) as defined in claim 29.
31. A compound of formula (B) according to claim 30 which is rapamycin 42-ester with 3-hydroxy-2-(2-iodo-acetoxymethyl)-2-methyl-propionic acid.
32. A compound of formula (B) according to claim 30 which is rapamycin 42-ester with 3-(2-iodo-acetoxy)-2-(2-iodo-acetoxymethyl)-2-methyl-propionic acid.
Applications Claiming Priority (3)
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| US23377600P | 2000-09-19 | 2000-09-19 | |
| US60/233,776 | 2000-09-19 | ||
| PCT/US2001/028575 WO2002024706A2 (en) | 2000-09-19 | 2001-09-13 | Water soluble rapamycin esters |
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| Publication Number | Publication Date |
|---|---|
| CA2421485A1 true CA2421485A1 (en) | 2002-03-28 |
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ID=22878646
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|---|---|---|---|
| CA002421485A Abandoned CA2421485A1 (en) | 2000-09-19 | 2001-09-13 | Water soluble rapamycin esters |
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| US (1) | US6432973B1 (en) |
| EP (1) | EP1319008B1 (en) |
| JP (1) | JP2004509898A (en) |
| CN (1) | CN1222529C (en) |
| AT (1) | ATE411321T1 (en) |
| AU (1) | AU2001290841A1 (en) |
| BR (1) | BR0113975A (en) |
| CA (1) | CA2421485A1 (en) |
| DE (1) | DE60136200D1 (en) |
| DK (1) | DK1319008T3 (en) |
| ES (1) | ES2313983T3 (en) |
| MX (1) | MXPA03002258A (en) |
| WO (1) | WO2002024706A2 (en) |
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| US5504091A (en) | 1993-04-23 | 1996-04-02 | American Home Products Corporation | Biotin esters of rapamycin |
| US5516770A (en) | 1993-09-30 | 1996-05-14 | American Home Products Corporation | Rapamycin formulation for IV injection |
| US5536729A (en) | 1993-09-30 | 1996-07-16 | American Home Products Corporation | Rapamycin formulations for oral administration |
| US5616588A (en) | 1993-09-30 | 1997-04-01 | American Home Products Corporation | Rapamycin formulation for IV injection |
| IL111004A (en) | 1993-09-30 | 1998-06-15 | American Home Prod | Oral rapamycin formulations |
| US5391730A (en) | 1993-10-08 | 1995-02-21 | American Home Products Corporation | Phosphorylcarbamates of rapamycin and oxime derivatives thereof |
| US5385908A (en) | 1993-11-22 | 1995-01-31 | American Home Products Corporation | Hindered esters of rapamycin |
| US5385909A (en) | 1993-11-22 | 1995-01-31 | American Home Products Corporation | Heterocyclic esters of rapamycin |
| US5385910A (en) | 1993-11-22 | 1995-01-31 | American Home Products Corporation | Gem-distributed esters of rapamycin |
| US5389639A (en) | 1993-12-29 | 1995-02-14 | American Home Products Company | Amino alkanoic esters of rapamycin |
| US5362718A (en) * | 1994-04-18 | 1994-11-08 | American Home Products Corporation | Rapamycin hydroxyesters |
| US5463048A (en) | 1994-06-14 | 1995-10-31 | American Home Products Corporation | Rapamycin amidino carbamates |
| US5491231A (en) | 1994-11-28 | 1996-02-13 | American Home Products Corporation | Hindered N-oxide esters of rapamycin |
| US5561138A (en) | 1994-12-13 | 1996-10-01 | American Home Products Corporation | Method of treating anemia |
| US5496832A (en) | 1995-03-09 | 1996-03-05 | American Home Products Corporation | Method of treating cardiac inflammatory disease |
| US5780462A (en) * | 1995-12-27 | 1998-07-14 | American Home Products Corporation | Water soluble rapamycin esters |
| US5989591A (en) | 1997-03-14 | 1999-11-23 | American Home Products Corporation | Rapamycin formulations for oral administration |
| US5985325A (en) | 1997-06-13 | 1999-11-16 | American Home Products Corporation | Rapamycin formulations for oral administration |
-
2001
- 2001-09-13 AT AT01970890T patent/ATE411321T1/en not_active IP Right Cessation
- 2001-09-13 JP JP2002529116A patent/JP2004509898A/en not_active Withdrawn
- 2001-09-13 CN CNB018159117A patent/CN1222529C/en not_active Expired - Fee Related
- 2001-09-13 EP EP01970890A patent/EP1319008B1/en not_active Expired - Lifetime
- 2001-09-13 DE DE60136200T patent/DE60136200D1/en not_active Expired - Fee Related
- 2001-09-13 MX MXPA03002258A patent/MXPA03002258A/en active IP Right Grant
- 2001-09-13 BR BR0113975-4A patent/BR0113975A/en not_active IP Right Cessation
- 2001-09-13 WO PCT/US2001/028575 patent/WO2002024706A2/en active Application Filing
- 2001-09-13 CA CA002421485A patent/CA2421485A1/en not_active Abandoned
- 2001-09-13 AU AU2001290841A patent/AU2001290841A1/en not_active Abandoned
- 2001-09-13 ES ES01970890T patent/ES2313983T3/en not_active Expired - Lifetime
- 2001-09-13 DK DK01970890T patent/DK1319008T3/en active
- 2001-09-18 US US09/954,782 patent/US6432973B1/en not_active Expired - Fee Related
Also Published As
| Publication number | Publication date |
|---|---|
| EP1319008B1 (en) | 2008-10-15 |
| WO2002024706A2 (en) | 2002-03-28 |
| DK1319008T3 (en) | 2009-02-09 |
| BR0113975A (en) | 2003-07-29 |
| JP2004509898A (en) | 2004-04-02 |
| CN1460108A (en) | 2003-12-03 |
| DE60136200D1 (en) | 2008-11-27 |
| ES2313983T3 (en) | 2009-03-16 |
| US6432973B1 (en) | 2002-08-13 |
| US20020055518A1 (en) | 2002-05-09 |
| EP1319008A2 (en) | 2003-06-18 |
| WO2002024706A3 (en) | 2002-10-10 |
| CN1222529C (en) | 2005-10-12 |
| MXPA03002258A (en) | 2003-06-24 |
| AU2001290841A1 (en) | 2002-04-02 |
| ATE411321T1 (en) | 2008-10-15 |
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Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| EEER | Examination request | ||
| FZDE | Discontinued |