Classifications

• • 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
• • 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
  • A61K47/54—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 compound
  • A61K47/55—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 compound the modifying agent being also a pharmacologically or therapeutically active agent, i.e. the entire conjugate being a codrug, i.e. a dimer, oligomer or polymer of pharmacologically or therapeutically active compounds
  • A61K47/552—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 compound the modifying agent being also a pharmacologically or therapeutically active agent, i.e. the entire conjugate being a codrug, i.e. a dimer, oligomer or polymer of pharmacologically or therapeutically active compounds one of the codrug's components being an antibiotic
• • 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
  • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
  • A61K31/47—Quinolines; Isoquinolines
  • A61K31/4738—Quinolines; Isoquinolines ortho- or peri-condensed with heterocyclic ring systems
  • A61K31/4745—Quinolines; Isoquinolines ortho- or peri-condensed with heterocyclic ring systems condensed with ring systems having nitrogen as a ring hetero atom, e.g. phenantrolines
• • 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
  • A61K47/54—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 compound
  • A61K47/55—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 compound the modifying agent being also a pharmacologically or therapeutically active agent, i.e. the entire conjugate being a codrug, i.e. a dimer, oligomer or polymer of pharmacologically or therapeutically active compounds
• • 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
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
  • A61P1/04—Drugs for disorders of the alimentary tract or the digestive system for ulcers, gastritis or reflux esophagitis, e.g. antacids, inhibitors of acid secretion, mucosal protectants
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
  • A61P1/18—Drugs for disorders of the alimentary tract or the digestive system for pancreatic disorders, e.g. pancreatic enzymes
• • A—HUMAN NECESSITIES
  • 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
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P15/00—Drugs for genital or sexual disorders; Contraceptives
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P15/00—Drugs for genital or sexual disorders; Contraceptives
  • A61P15/14—Drugs for genital or sexual disorders; Contraceptives for lactation disorders, e.g. galactorrhoea
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P3/00—Drugs for disorders of the metabolism
  • A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
  • A61P3/10—Drugs for disorders of the metabolism for glucose homeostasis for hyperglycaemia, e.g. antidiabetics
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P35/00—Antineoplastic agents
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P35/00—Antineoplastic agents
  • A61P35/02—Antineoplastic agents specific for leukemia
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P35/00—Antineoplastic agents
  • A61P35/04—Antineoplastic agents specific for metastasis

Definitions

• the present invention relates to a method of treating resistant or refractory cancers.
• the invention relates to a method of treating cancers resistant or refractory to camptothecin or CPT-11 using polyethylene glycol conjugates of 7-ethyl-10- hydroxycamptothecin.
• anti-cancer agents have been developed in efforts to treat cancers. Many of those potential anti-cancer agents have unfortunately shown drug resistance tlirough a variety of mechanisms. Some tumors do not respond to certain types of anti-cancer agents after initial short therapeutic responses are shown. In some cases, tumor shrinkage reverses and tumors start to grow again in spite of the cancer initially responding to anti-cancer agents.
• camptothecin One potent anti-cancer agent is camptothecin. Camptothecin and related analogs are known as DNA topoisomerase I inhibitors.
• Irinotecan CPT-11, Camptosar®
• CPT-11, Camptosar® an active metabolite of CPT-11, 7-ethyl-l O-hydroxycamptothecin, is thought to also have some anticancer activity.
• drug resistance has been observed with the use of camptothecin and camptothecin derivatives. For example, resistance to 9-amino or 9-nitro substituted camptothecins has been reported in common cancers. See US Patent No. 6,194,579.
• a method of treating a resistant or refractory cancer in a mammal including: administering an effective amount of a compound of formula (I):
• R ⁇ , R 2 , R 3 and R 4 are independently OH or
• L is a bifunctional linker; in is 0 or a positive integer; and n is a positive integer; provided that Ri, R 2 , R 3 and R 4 are not all OH; or a pharmaceutically acceptable salt thereof to the mammal.
• the polymeric prodrugs of 7-ethyl-10- hydroxycamptothecin for treatment of the resistant or refractory cancer employ four-arm PEG-7-ethyl-lO-hydroxycamptothecin conjugates having the structure of
• n is from about 28 to about 341, preferably from about 1 14 to about 227, and more preferably about 227.
• the resistant or refractory cancers which can be treated with the methods described herein include solid tumors, lymphomas, lung cancer, small cell lung cancer, acute lymphocytic leukemia (ALL), breast cancer, colorectal cancer, pancreatic cancer, glioblastoma, ovarian cancer and gastric cancer.
• ALL acute lymphocytic leukemia
• breast cancer colorectal cancer
• pancreatic cancer pancreatic cancer
• glioblastoma ovarian cancer
• gastric cancer gastric cancer.
• One aspect of the invention provides the method of treating cancers resistant or refractory to chemotherapy.
• the treatment is effective for cancers resistant or refractory to camptothecin (CPT) or CPT-1 1 associated therapy.
• CPT camptothecin
• the present invention provides a method of treating cancers showing topoisomerase 1 mediated resistance or refractory phenomenon.
• the present invention provides a method of treating cancers resistant or refractory to therapies associated with administration of polymeric prodrug forms of CPT or CPT-1 1 such as polyethylene glycol conjugates of CPT or CPT-11.
• the polymeric prodrugs of 7-ethyl-10-hydroxycamptothecin according to the present invention are effective to cancers resistant or refractory at the onset of treatment or subsequent round therapies.
• the present invention allows treatment of refractory cancers that are sensitive to CPT-11, i.e. which appear to be inhibited in the first round treatment but become resistant to in the second or subsequent rounds of therapies.
• the polymeric prodrugs of 7-ethyl-lO-hydroxycamptothecin can be further effective for treatment of recurring cancers after treatment is discontinued.
• the polymeric prodrugs of 7-ethyl-10-hydroxy- camptothecin are administered in amounts of from about 0.1 to about 45 mg/m7dose based on the non-polymer portion of the conjugate.
• the polymeric prodrugs described herein are administered once every three weeks for each treatment cycle or once weekly for three weeks, followed by one week rest period for each cycle until the desired results are observed.
• One advantage of the present invention is that patients can be treated concurrently or sequentially with an effective amount of the polymeric prodrugs of 7-ethyl-10- hydroxy camp tothecin in combination with another anti-cancer therapeutic agent for synergistic benefit.
• the term “residue” shall be understood to mean that portion of a compound, to which it refers, i.e. 7-ethyl-10-hydroxycamptothecin, amino acid, etc. that remains after it has undergone a substitution reaction with another compound.
• PEG residue shall each be understood to mean that portion of the polymer or PEG which remains after it has undergone a reaction with 7-ethyl-l O-hydroxycamptothecin-containing compounds.
• alkyl refers to a saturated aliphatic hydrocarbon, including straight-chain, branched-chain, and cyclic alkyl groups.
• alkyl also includes alkyl -thio-alkyl, alkoxyalkyl, cycloalkylalkyl, heterocycloalkyl, Ci -6 hydrocarbonyl, groups.
• the alkyl group has 1 to 12 carbons. More preferably, it is a lower alkyl of from about 1 to 7 carbons, yet more preferably about 1 to 4 carbons.
• the alkyl group can be substituted or unsubstituted.
• the substituted group(s) preferably include halo, oxy, azido, nitro, cyano, alkyl, alkoxy, alkyl - thio, alkyl-thio-alkyl, alkoxyalkyl, alkylamino, trihalomethyl, hydroxyl, mercapto, hydroxy, cyano, alkylsilyl, cycloalkyl, cycloalkylalkyl, heterocycloalkyl, heteroaryl, alkenyl, alkynyl, C i- 6 hydrocarbonyl, aryl, and amino groups.
• substituted refers to adding or replacing one or more atoms contained within a functional group or compound with one of the moieties from the group of halo, oxy, azido, nitro, cyano, alkyl, alkoxy, alkyl- thio, alkyl-thio-alkyl, alkoxyalkyl, alkylamino, trihalomethyl, hydroxyl, raercapto, hydroxy, cyano, alkylsilyl, cycloalkyl, cycloalkylalkyl, heterocycloalkyl, heteroaryl, alkenyl, alkynyl, C i- 6 hydrocarbonyl, aryl, and amino groups.
• alkenyl refers to groups containing at least one carbon- carbon double bond, including straight-chain, branched-chain, and cyclic groups.
• the alkenyl group has about 2 to 12 carbons. More preferably, it is a lower alkenyl of from about 2 to 7 carbons, yet more preferably about 2 to 4 carbons.
• the alkenyl group can be substituted or unsubstituted. When substituted the substituted group(s) preferably include halo, oxy.
• alkylamino trihalomethyl, hydroxyl, mercapto, hydroxy, cyano, alkylsilyl, cycloalkyl, cycloalkylalkyl, heterocycloalkyl, heteroaryl, alkenyl, alkynyl, Ci -6 hydrocarbonyl, aryl, and amino groups.
• alkynyl refers to groups containing at least one carbon- carbon triple bond, including straight-chain, branched-chain, and cyclic groups.
• the alkynyl group has about 2 to 12 carbons. More preferably, it is a lower alkynyl of from about 2 to 7 carbons, yet more preferably about 2 to 4 carbons.
• the alkynyl group can be substituted or unsubstituted.
• the substituted group(s) preferably include halo, oxy, azido, nitro, cyano, alkyl, alkoxy, alkyl-thio, alkyl-thio-alkyl, alkoxyalkyl, alkylamino, trihalomethyl, hydroxy!, mercapto, hydroxy, cyano, alkylsilyl, cycloalkyl, cycloalkylalkyl, heterocycloalkyl, heteroaryl, alkenyl, alkynyl, Cj -6 hydrocarbonyl, aryl, and amino groups.
• alkynyl include propargyl, propyne, and 3-hexyne.
• aryl refers to an aromatic hydrocarbon ring system containing at least one aromatic ring.
• the aromatic ring can optionally be fused or otherwise attached to other aromatic hydrocarbon rings or non-aromatic hydrocarbon rings.
• aryl groups include, for example, phenyl, naphthyl, 1 ,2,3,4-tetrahydronaphthalene and biphenyl.
• Preferred examples of aryl groups include phenyl and naphthyl.
• cycloalkyl refers to a Cj -8 cyclic hydrocarbon.
• examples of cycloalkyl include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl and cyclooctyl.
• cycloalkenyl refers to a C 3- g cyclic hydrocarbon containing at least one carbon-carbon double bond.
• examples of cycloalkenyl include cyclopentenyl, cyclopentadienyl, cyclohexenyl, 1,3-cyclohexadienyl, cycloheptenyl, cycloheptatrienyl, and cyclooctenyl.
• cycloalkylalkyl refers to an alklyl group substituted with a C 3-S cycloalkyl group.
• examples of cycloalkylalkyl groups include cyclopropylmethyl and cyclopentylethyl.
• alkoxy refers to an alkyl group of indicated number of carbon atoms attached to the parent molecular moiety through an oxygen bridge.
• alkoxy groups include, for example, methoxy, ethoxy, propoxy and isopropoxy.
• alkylaryl refers to an aryl group substituted with an alkyl group.
• aralkyl group refers to an alkyl group substituted with an aryl group.
• alkoxyalkyl refers to an alkyl group substituted with an alkloxy group
• alkyl-thio-alkyl refers to an alkyl-S-alkyl thioether, for example methyl thiomethyl or methylthioethyl.
• amino refers to a nitrogen containing group as is known in the art derived from ammonia by the replacement of one or more hydrogen radicals by organic radicals.
• acylamino and “alkyl amino” refer to specific N- substituted organic radicals with acyl and alkyl substituent groups respectively.
• alkylcarbonyl refers to a carbonyl group substituted with alkyl group.
• halogen' or halo refer to fluorine, chlorine, bromine, and iodine.
• heterocycloalkyl refers to a non-aromatic ring system containing at least one heteroatom selected from nitrogen, oxygen, and sulfur.
• the heterocycloalkyl ring can be optionally fused to or otherwise attached to other heterocycloalkyl rings and/or non-aromatic hydrocarbon rings.
• Preferred heterocycloalkyl groups have from 3 to 7 members. Examples of heterocycloalkyl groups include, for example, piperazine, morpholine, piperidine, tetrahydrofuran, pyrrolidine, and pyrazole.
• Preferred heterocycloalkyl groups include piperidinyl, piperazinyl, morpholinyl, and pyrolidinyl.
• heteroaryl refers to an aromatic ring system containing at least one heteroatom selected from nitrogen, oxygen, and sulfur.
• the heteroaryl ring can be fused or otherwise attached to one or more heteroaryl rings, aromatic or non-aromatic hydrocarbon rings or heterocycloalkyl rings.
• heteroaryl groups include, for example, pyridine, furan, thiophene, 5,6,7,8-tetrahydroisoquinoline and pyrimidine.
• heteroaryl groups include thienyl, benzothienyl, pyridyl, quinolyl, pyrazinyl, pyrimidyl, imidazolyl, benzimidazolyl, furanyl, benzofuranyl, thiazolyl, benzothiazolyl, isoxazolyl, oxadiazolyl, isothiazolyl, benzisothiazolyl, triazolyl, tetrazolyl, pyrrolyl, indolyl, pyrazolyl, and benzopyrazolyl.
• heteroatom refers to nitrogen, oxygen, and sulfur.
• substituted alkyls include carboxyalkyls, aminoalkyls, dialkylaminos, hydroxyalkyls and mercaptoalkyls; substituted alkenyls include carboxyalkenyls, aminoalkenyls, dialkenyl aminos, hydroxyalkenyls and mercaptoalkenyls; substituted alkynyls include carboxyalkynyls, aminoalkynyls, dialkynyl aminos, hydroxyalkynyls and mercaptoalkynyls; substituted cycloalkyls include moieties such as 4-chlorocyclohexyl; aryls include moieties such as napthyl; substituted aryls include moieties such as 3-bromo phenyl; aralkyls include moieties such as tolyl; heteroalkyls include moieties such as ethylthiophene; substituted heteroalkyls
• positive integer shall be understood to include an integer equal to or greater than 1 and as will be understood by those of ordinary skill to be within the realm of reasonableness by the artisan of ordinary skill.
• FIG. 1 shows anticancer activity of four-arm PEG-Gly-7-ethyl-10- hydroxycamptothecin in treatment of CPT-] 1 refractory colorectal tumor as described in Example 1.
• FIG. 2 shows anticancer activity of four-a ⁇ n PEG-Gly-7-ethyl-10- hydroxycamptothecin in treatment of CPT-1 1 refractory colorectal tumor as described in Example 2.
• FIG. 3 shows in vitro cytotoxicity of four-a ⁇ n PEG-Gly-7-ethyl-l 0- hydroxycamptotehcin in the cells refractory to CPT as described in Example 3.
• FIG. 4 shows in vitro cytotoxicity of four-arm PEG-Gly-7-ethyl-lO- hydroxycamptotehcin in the cells non-refractory to CPT as described in Example 3.
• methods of treating a resistant or refractory cancer in a mammal comprising: administering an effective amount of a compound of formula (I): wherein
• Ri, R 2 , R 3 and R 4 are independently OH or
• L is a bifunctional linker; m is 0 or a positive integer, preferably 1 ; and n is a positive integer; provided that R 1 , R 2 , R 3 and R 4 are not all OH; or a pharmaceutically acceptable salt thereof to the mammal in need thereof.
• one, two or three Of R 1 , R 2 , R 3 and R 4 can be CH 3 .
• refractory or resistant cancers are defined as cancers that do not respond to previous anticancer therapy or treatment.
• the cancers are refractory or resistant to CPT-11 treatment.
• the cancers can be resistant or refractory at the beginning of treatment, or they may become resistant or refractory during treatment.
• Refractory cancers include tumors that do not respond at the onset of treatment or respond initially for a short period but fail to respond to treatment.
• Refractory cancers also include tumors that respond to treatment with anticancer therapy but fail to respond to subsequent rounds of therapies.
• refractory cancers also encompass tumors that appear to be inhibited by treatment with anticancer therapy but recur up to five years, sometimes up to ten years or longer after treatment is discontinued.
• the anticancer therapy can employ chemotherapeutic agents alone, radiation alone or combinations thereof.
• chemotherapeutic agents alone, radiation alone or combinations thereof.
• the refractory cancers are interchangeable with resistant cancers.
• successful treatment of a resistant or refractory cancer shall be understood to mean that resistant or refractory symptoms or conditions are prevented, minimized or attenuated during and/or after anticancer treatment, when compared to mat observed in the absence of the treatment described herein.
• the minimized, attenuated or prevented refractory conditions can be confirmed by clinical markers contemplated by the artisan in the field.
• successful treatment of refractory or resistant cancer shall be deemed to occur when at least 5 % or preferably 10%, more preferably 20% or higher (i.e., 30, 40, 50 % or more) inhibition or decrease in tumor growth and/or recurrence including other clinical markers contemplated by the artisan in the field is realized when compared to that observed in the absence of the treatment described herein.
• the resistant or refractory cancers can be one or more of the following: solid tumors, lymphomas, small cell lung cancer, acute lymphocytic leukemia (ALL), pancreatic cancer, glioblastoma, ovarian cancer, gastric cancers, etc.
• the methods are useful for, among other things, treating neoplastic disease, reducing tumor burden, preventing metastasis of neoplasms and preventing recurrences of tumor/neoplastic growths in mammals.
• the resistant or refractory cancers are solid tumors or metastatic cancers.
• the resistant or refractory cancer is colorectal cancer.
• the present invention provides methods of treating resistant or refractory cancers to chemotherapy.
• the present invention provides methods of treating cancers which are resistant or refractory to camptothecin (CPT) or camptothecin analog therapy.
• CPT camptothecin
• the methods described herein can be effective to treat cancers resistant or refractory to CPT or CPT analog conjugated to polymers such as polyethylene glycol.
• the present invention provides methods of treating cancers which are resistant or refractory to camptothecin or CPT-1 1 therapy. Camptothecin and certain related analogs share the structure:
• the A ring in either or both of the 10- and 1 1 -positions can be substituted with an OH.
• the A ring can also be substituted with a straight or branched Ci -3O alkyl or C M 7 alkoxy, optionally linked to the ring by a heteroatom i.e.- O or -S.
• the B ring can be substituted in the 7-position with a straight or branched Cj -3 O alkyl (preferably C 2 alkyl), Cs -S cycloakyl, C] -3 O alkoxy, phenyl alkyl, etc., alkyl carbamate, alkyl carbazides, phenyl hydrazine derivatives, etc.
• Cj -3 O alkyl preferably C 2 alkyl
• Cs -S cycloakyl C] -3 O alkoxy
• phenyl alkyl, etc. alkyl carbamate, alkyl carbazides, phenyl hydrazine derivatives, etc.
• Other substitutions are possible in the C 5 D and E rings. See, for example, U.S. Patent Nos. 5,004,758; 4,943,579; 4,473,692; RE32,518, the contents of which are incorporated herein by reference.
• the 10-hydroxycamptothecin, 11 -hydroxycamptothecin and the 10,1 1 -dihydroxycamtothecin analogs occur naturally as one of the minor components in C. Acuminata and its relatives. Additional substitutions to these compounds, i.e. 7-alkyl-, 7-substituted alkyl-, 7-amino-, 7-aminoalkyl-, 7-aralkyl-, 9-alkyl- s 9-aralkyl- camptothecin etc. derviatives are made using known synthetic techniques.
• Some camptotheca alkaloids have the structure shown below:
• R 7 is one OfNO 2 , NH 2 , N 3 , hydrogen, halogen (F, Cl, Br, I), COOH, OH, 0-C 1 -8 alkyl, SH, S-C -3 alkyl, CN, CH 2 NH 2 , NH-C 1-3 alkyl, CH 2 -NH-C 1-3 alkyl, N(C 1 -3 alkyl) 2 , CH 2 N(C N3 alkyl), O-, NH- and S-CH 2 CH 2 N(CH 2 CH 2 OH) 2 ,, O- > NH- and S-CH 2 CH 2 CH 2 N(CH 2 CH 2 OH) 2 , O-, NH- and
• R 9 and Ri 0 are, independently, hydrogen, C
• R 9 can be hydrogen, C !-6 alkyl, C 3-7 cycloalkyl, C 3-7 cycloalkyl-C 1-6 alkyl, C 2 -6 alkenyl, hydroxy-Ci -6 alkyl, Ci -6 alkoxy-C
• R 9 and R J Q taken together with the nitrogen atom to which they are attached form a saturated 3-7 membered heterocyclic ring which may contain a O, S or NRj2 group, where R [2 is hydrogen, C
• Ri 3 is hydrogen, C 1-6 alkyl, perhalo-C 1-6 alkyl, Ci-6 alkoxy, aryl, and aryl substituted with one or more of Ci -6 alkyl, perhalo-Ci-e alkyl, hydroxy-Ci. 6 alkyl, or Ci -6 alkoxy-C t-6 alkyl groups;
• Ri io-R ⁇ 11 are each independently selected from among hydrogen; halo; acyl; alkyl
• the aryl groups can be phenyl and naphthyl.
• Suitable heterocyclic rings when R 9 and R 1 O are taken together with the nitrogen atom to which they are attached include: aziridine, azetidine, pyrrolidine, piperidine, hexamethylenimine, imidazolidine, pyrazolidine, isoxazolidine, piperazine, N-methylpiperazine, tetrahydroazepine, N-methyl- tetrahydroazepine, thiazolidine, etc.
• the treatment of the present invention includes administering an effective amount of the compounds described herein to a mammal with resistant or refractory cancers showing topoisomerase I mediated resistance or refractory phenomenon.
• the present invention provides methods of treating resistant or refractory cancers associated with radiation therapy alone or radiation therapy in combination with a second chemotherapy.
• Standard protocols of radiation therapy are well known in the art and thus, the combination therapy using the compounds described herein can be done without undue experimentation.
• the treatment of the present invention includes administering an effective amount of the compounds described herein alone or in combination, simultaneously or sequentially, with a second chemotherapeutic agent.
• the multi-arm polymeric prodrugs of 7-ethyl-l O-hydroxycamptothecin can be administered concurrently with the chemotherapeutic agent or after the administration of the chemotherapeutic agent.
• the compounds employed in the present invention can be administered during or after treatment of the second chemotherapeutic agent.
• a non-liming list of the second chemotherapeutic agents includes:
• DNA topoisomerase inhibitor adriamycin, amsacrine, camptothecin, CPT-1 1, daunorubicin, dactinomycin, doxorubicin, eniposide, epirubicin, etoposide, idarubicin, or mitoxantrone;
• microtubule inhibiting drug such as a taxane, including paclitaxel, docetaxel, vincristin, vinblastin, nocodazole, epothilones and navelbine;
• DNA damaging agent actinomycin, amsacrine, anthracyclines, bleomycin, busulfan, camptothecin, carboplatin, chlorambucil, cisplatin, cyclophosphamide, Cytoxan, dactinomycin, daunorubicin, docetaxel, doxorubicin, epirubicin, hexamethylmelamineoxaliplatin, iphosphamide, melphalan, merchlorehtamine, mitomycin, mitoxantrone, nitrosourea, plicamycin, procarbazine, taxol, taxotere, teniposide, triethylenethiophosphoramide or etoposide (VP 16);
• antimetabolite folate antagonist
• nucleoside analog 5-fluorouracil; cytosine arabinoside, azacitidine, 6- mercaptopurine, azathioprine; 5- ⁇ odo-2'-deoxyuridine; 6-tliioguanine, 2-deoxycoforaiycin, cladribine, cytarabine, fiudarabine, mercaptopurine, thioguanine, pentostatin, AZT (zidovudine), ACV, valacylovir, famiciclovir, acyclovir, cidofovir, penciclovir, ganciclovir, Ribavirin, ddC, ddl (zalcitabine), lamuvidine, Abacavir, Adefovir, Didanosine, d4T (stavudine), 3TC, BW 1592, PMEA/bis-POM PME
• Other potential anti-cancer agents are selected from altretamine, aminoglutethimide, amsacrine, anastrozole, asparaginase, beg, bicalutamide, bleomycin, buserelin, busulfan, calcium folinate, campotliecin, capecitabine, carboplatin, carmustine, chlorambucil, cisplatin, cladribine, clodronate, colchicine, crisantaspase, cyclophosphamide, cyproterone, cytarabine, dacarbazine, dactinomycin, daunorubicin, dienestrol, diethylstilbestrol, docetaxel, doxorubicin, epirubicin, estradiol, estramustine, etoposide, exemestane, filgrastim, fludarabine, fludrocortisone, fluorouracil, fluoxymesterone, flutamide,
• the treatment of resistant or refractory cancers uses the compounds among:
• One particularly preferred compound is
• n is from about 28 to about 341 , preferably from about 1 14 to about 227, or more preferably about 227.
• One preferred embodiment (compound 9) of the present invention has a molecular weight of about 40,000 da and have the structure of:
• the unexpected efficacy of the methods described herein in treatment of CPT-1 1 refractory tumors can be attributed, at least in part, to the favorable pharmacokinetic and biodistribution properties of the polymeric compounds described herein.
• the unexpected efficacy of the compounds described herein can also be based in part on a novel mechanism of action for the drug in vivo. It has been reported that topotecan, another TOPl inhibitor, inhibits hypoxia-inducible factor (HIF)-I a, leading to marked decrease of angiogenesis and significant tumor growth inhibition.
• HIF hypoxia-inducible factor
• inventive treatment induces a decrease in HIF-I ⁇ in cells, which then accumulate the compounds described herein due to an EPR effect in CPT-1 1 refractory (or sensitive) tumors.
• CPT-1 1 fails to induce a decrease in HIF-I ⁇ in CPT-1 1 refractory tumors, leading to even more angiogenesis.
• treatment of highly vascular tumors can benefit from accumulation of the inventive compounds described herein due to enhanced EPR effects.
• CPT-11 resistant tumors may have lower levels of TOPl, since low levels of TOPl have been linked to CPT-11 resistance in tissue culture.
• the polymeric ester derivatives of 7-ethyl-lO-hyroxycamptothecin according to the therapy described herein can also provide higher exposure of 7-ethyl-lO-hyroxycarnptothecin to cells in vivo than CPT-1 1. Drug concentrations can be sufficient to kill cells even with low levels of TOPl.
• variable levels of carboxyl esterase can be another contributing factor to CPT-1 1 resistance, and this enzyme is not required for release of 7-ethyl-l O- hyroxycamptothecin from the polymeric ester derivatives of 7-ethyl-10-hyroxycamptothecin conjugates described herein.
• carboxyl esterase can be another contributing factor to CPT-1 1 resistance, and this enzyme is not required for release of 7-ethyl-l O- hyroxycamptothecin from the polymeric ester derivatives of 7-ethyl-10-hyroxycamptothecin conjugates described herein.
• the polymeric prodrugs of 7-ethyl-lO-hydroxycaraptothecin include four-arm PEG attached to 20-OH group of 7-ethyl-lO-hydroxycamptothecin through a bifunctional linker.
• the polymeric prodrugs of 7-ethyl-lO-hydroxy- camptothecin include four-arm PEG, prior to conjugation, having the following structure of
• n is a positive integer
• the polymers are those described in NOF Corp. Drug Delivery System catalog, Ver. 8, April
• the degree of polymerization for the polymer (n) is from about 28 to about 341 to provide polymers having a total molecular weight of from about 5,000 Da to about 60,000 Da, and preferably from about 1 14 to about
• n represents the number of repeating units in the polymer chain and is dependent on the molecular weight of the polymer, In one particularly preferred embodiment of the invention, n is about 227 to provide the polymeric portion having a total molecular weight of about 40,000 Da.
• L is a residue of an amino acid.
• the amino acid can be selected from any of the known naturally-occurring L-amino acids, e.g., alanine, valine, leucine, isoleucine, glycine, serine, threonine, methionine, cysteine, phenylalanine, tyrosine, tryptophan, aspartic acid, glutamic acid, lysine, arginine, histidine, proline, and/or a combination thereof, to name but a few.
• L can be a peptide residue.
• the peptide can range in size, for instance, from about 2 to about 10 amino acid residues.
• amino acid analogs and derivates include:
• one arm of the four-arm PEG is shown.
• One arm, up to four a ⁇ ns of the four-ami PEG can be conjugated with 7-ethyl-10-hydroxy- camptothecin.
• compounds of the present invention include a glycine residue as the linker group (L).
• L after attachment between the camptothecin analog and polymer is selected among:
• R21-R 29 are independently selected among hydrogen, amino, substituted amino, azido, carboxy, cyano, halo, hydroxyl, nitro, silyl ether, sulfonyl, mercapto, Ci -6 alkylmercapto, arylmercapto, substituted arylmercapto, substituted
• alkanoyl arylcarbonyl, C 2-6 alkoxycarbonyl, aryloxycarbonyl, C 2-6 alkanoyloxy, arylcarbonyloxy, C 2-6 substituted alkanoyl, substituted arylcarbonyl, C 2 . 6 substituted alkanoyloxy, substituted aryloxycarbonyl, C 2-6 substituted alkanoyloxy, substituted and arylcarbonyloxy;
• (t), (V) and (y) are independently selected from zero or a positive integer, preferably from about 1 to about 10;
• L can include:
• the compounds include from 1 to about 10 units of the bifunctional linker. In some preferred aspects of the present invention, the compounds include one unit of the bifunctional linker and thus m is 1.
• the polymeric 7-ethyl-lO-hydroxycamptothecin prodrugs described herein are prepared by reacting one or more equivalents of an activated multi-arm polymer with, for example, one or more equivalents per active site of amino acid-(20)-7-ethyl- 1 O- hydroxycamptothecin compound under conditions which are sufficient to effectively cause the amino group to undergo a reaction with the carboxylic acid of the polymer and form a linkage. Details of the synthesis are described in US Patent Application No.
• compositions containing the polymer conjugates of the present invention may be manufactured by processes well known in the art, e.g., using a variety of well-known mixing, dissolving, granulating, levigating, emulsifying, encapsulating, entrapping or lyophilizing processes.
• the compositions may be formulated in conjunction with one or more physiologically acceptable carriers comprising excipients and auxiliaries which facilitate processing of the active compounds into preparations which can be used pharmaceutically. Proper formulation is dependent upon the route of administration chosen. Parenteral routes are preferred in many aspects of the invention.
• the compounds of the invention may be formulated in aqueous solutions, preferably in physiologically compatible buffers such as physiological saline buffer or polar solvents including, without limitation, a pyrrolidone or dimethylsulfoxide.
• physiologically compatible buffers such as physiological saline buffer or polar solvents including, without limitation, a pyrrolidone or dimethylsulfoxide.
• the compounds are preferably formulated for parenteral administration, e.g., by bolus injection or continuous infusion.
• Formulations for injection may be presented in unit dosage form, e.g.j in ampoules or in multi-dose containers.
• Useful compositions include, without limitation, suspensions, solutions or emulsions in oily or aqueous vehicles, and may contain adjuncts such as suspending, stabilizing and/or dispersing agents.
• Pharmaceutical compositions for parenteral administration include aqueous solutions of a water soluble form, such as, without limitation, a salt of the active compound. Additionally, suspensions of the active compounds may be prepared in a lipophilic vehicle.
• Suitable lipophilic vehicles include fatty oils such as sesame oil, synthetic fatty acid esters such as ethyl oleate and triglycerides, or materials such as liposomes.
• Aqueous injection suspensions may contain substances that increase the viscosity of the suspension, such as sodium carboxym ethyl cellulose, sorbitol, or dextran.
• the suspension may also contain suitable stabilizers and/or agents that increase the solubility of the compounds to allow for the preparation of highly concentrated solutions.
• the active ingredient may be in powder form for constitution with a suitable vehicle, e.g., sterile, pyrogen-free water, before use.
• the compounds can be formulated by combining the active compounds with pharmaceutically acceptable carriers well-known in the art.
• Such carriers enable the compounds of the invention to be formulated as tablets, pills, lozenges, dragees, capsules, liquids, gels, syrups, pastes, slurries, solutions, suspensions, concentrated solutions and suspensions for diluting in the drinking water of a patient, premixes for dilution in the feed of a patient, and the like, for oral ingestion by a patient.
• Pharmaceutical preparations for oral use can be made using a solid excipient, optionally grinding the resulting mixture, and processing the mixture of granules, after adding other suitable auxiliaries if desired, to obtain tablets or dragee cores.
• Useful excipients are, in particular, fillers such as sugars, including lactose, sucrose, mannitol, or sorbitol, cellulose preparations such as, for example, maize starch, wheat starch, rice starch and potato starch and other materials such as gelatin, gum tragacanth, methyl cellulose, hydroxypropyl- methylcellulose, sodium carboxy- methylcellulose, and/or polyvinylpyrrolidone (PVP).
• disintegrating agents may be added, such as cross-linked polyvinyl pyrrolidone, agar, or alginic acid. A salt such as sodium alginate may also be used.
• the compounds of the present invention can conveniently be delivered in the form of an aerosol spray using a pressurized pack or a nebulizer and a suitable propellant
• the compounds may also be formulated in rectal compositions such as suppositories or retention enemas, using, e.g., conventional suppository bases such as cocoa butter or other glycerides.
• the compounds may also be formulated as depot preparations. Such long acting formulations may be administered by implantation (for example, subcutaneously or intramuscularly) or by intramuscular injection.
• the compounds of this invention may be formulated for this route of administration with suitable polymeric or hydrophobic materials (for instance, in an emulsion with a pharmacologically acceptable oil), with ion exchange resins, or as a sparingly soluble derivative such as, without limitation, a sparingly soluble salt.
• the compounds may be delivered using a sustained-release system, such as semi-permeable matrices of solid hydrophobic polymers containing the therapeutic agent.
• sustained-release materials have been established and are well known by those skilled in the art.
• Sustained-release capsules may, depending on their chemical nature, release the compounds for a few weeks up to over 100 days. Depending on the chemical nature and the biological stability of the particular compound, additional stabilization strategies may be employed.
• a therapeutically effective amount refers to an amount of compound effective to prevent, alleviate or ameliorate the resistance or refractory phenomenon to anti-cancer agents such as camptothecin or related analog, for example, CPT-1 1. Determination of a therapeutically effective amount is well within the capability of those skilled in the art, especially in light of the disclosure herein.
• the therapeutically effective amount can be estimated initially from in vitro assays. Then, the dosage can be formulated for use in animal models so as to achieve a circulating concentration range that includes the effective dosage. Such information can then be used to more accurately determine dosages useful in patients.
• the amount of the composition, e.g., used as a prodrug, that is administered will depend upon the parent molecule included therein. Generally, the amount of prodrug used in the treatment methods is that amount which effectively achieves the desired therapeutic result in mammals. Naturally, the dosages of the various prodrug compounds can vary somewhat depending upon the parent compound, rate of in vivo hydrolysis, molecular weight of the polymer, etc. In addition, the dosage, of course, can vary depending upon the dosage form and route of administration.
• polymeric ester derivatives of 7-ethyl-10- hyroxycamptothecin described herein can be administered in amounts ranging from about 0.1 to about 30 mg/kg/dose and preferably about 0.2 to about 10 mg/kg/dose, yet preferably from about 0.6 to about 6 mg/kg/dose for systemic delivery.
• the treatment of the present invention includes administering the compounds described herein in an amount of from about 0.3 to about 6 mg/kg/dose to a mammal having resistant or refractory cancers to such as CPT and CPT-1 1 therapies.
• the amounts of the compounds administered can be based on body surface of human or other mammals.
• the treatment of the present invention includes administering the compounds described herein in an amount of from about 0.1 to about 45 mg/m 2 body surface/dose.
• the amounts of the compounds described herein range from about 0.2 to about 25 mg/m 2 body surface/dose.
• Some preferred doses include one of the following: 1.25, 2.0 2.5, 3.3, 5, 10, and 16.5 mg/m 2 /dose.
• the amounts administered can range from about 1.25 to about 16.5 mg/m 2 body surface/dose. Alternatively, they can be from about 2.5 to about 13 mg/m 2 body surface/dose or from about 2 to about 5 mg/m 2 body surface/dose.
• the treatment protocol can be based on a single dose administered once every three weeks or divided into multiple doses which are given as part of a multi-week treatment protocol.
• the treatment regimens can include one dose every three weeks for each treatment cycle and, alternatively one dose weekly for three weeks followed by one week off for each cycle.
• the precise dose will depend on the stage and severity of the condition, and the individual characteristics of the patient being treated, as will be appreciated by one of ordinary skill in the art. It is also contemplated that the treatment continues until satisfactory results are observed, which can be as soon as after 1 cycle although from about 3 to about 6 cycles or more cycles may be required.
• the treatment protocol includes administering the amount ranging from about 1.25 to about 16.5 mg/m 2 body surface/dose every three weeks repeating for about 3 cycles or more.
• the amount administered per each cycle can range more preferably from about 2.5 to about 16.5 mg/m 2 body surface/dose.
• the compounds described herein can be administered weekly for three weeks, followed by one week without treatment and repeating for about 3 cycles or more until the desired results are observed.
• the polymeric ester derivatives of 7-ethyl-10- hyroxycamptothecin can be administered one dose such as 10 mg/m 2 every three weeks in treatment of colon cancer.
• the dosage of treatment cycle can be designed as an escalating dose regimen when two or more treatment cycles are applied.
• the polymeric drug is preferably administered via IV infusion.
• the dosage amount mentioned is based on the amount of 7-ethyI-l O-hydroxycamptothecin rather than the amount of polymeric conjugate administered. It is contemplated that the treatment will be given for one or more cycles until the desired clinical result is obtained.
• the exact amount, frequency and period of administration of the compound of the present invention will vary, of course, depending upon the sex, age and medical condition of the patient as well as the severity of the disease as determined by the attending clinician.
• Still further aspects include combining the therapy described herein with other anticancer therapies for synergistic or additive benefit.
• the compounds described herein can be administered in combination with Erbitux® (cetuximab). 400 mg/m 2 Erbitux® plus the compounds described herein can be administered as an initial dose followed by 250 mg/m weekly until disease progresses. Details of Erbitux® dosage information are described in the package insert, the contents of which are incorporated herein.
• mice Therapeutic efficacy of four-arm PET-Gly-(7 ⁇ ethyl-10-hydroxycamptothecin) against a refractory human HT-29 colorectal tumor grown in nude mice was determined.
• Human HT-29 colorectal tumors were established in nude mice by subcutaneous injection of 1 x 10 6 cells/mouse into a right auxiliary flank. When tumors reached an average volume of 100 mm 3 , mice were treated with CPT-1 1 (40 mg/kg/dose; q2d x 4). Mice were monitored for tumor growth. On day 15, mice with tumors that did not respond to CPT-1 1 therapy (tumor volume > 3 x initial tumor volume at the start of CPT-1 1 therapy) were considered CPT-1 1 refractory.
• mice were selected, randomized and divided into two groups. One group was treated with MTD of CPT-11 (40mg/kg/dose; q2d x 5) and the other group was treated with MTD of four-arm 40K PEG-Gly-(7-ethy ⁇ -10-hydroxycamptothecm) (compound 9) (10 mg/kg/dose q2d x 5) starting day 16.
• the drugs were administered intravenously via the tail vein.
• Fig. 1 Tumors continued to grow in the CPT-1 1 refractory mice further treated with CPT-1 1. On day 42, tumor volume increased by 255% compared to day 15. In the mice treated with four-arm 4(X PEG-Gly-(7-ethyl-10-hydroxy- camptothecin) (compound 9), tumor volume decreased by 25% compared to day 15 on day 42. 29% and 100% of animals treated with CPT-11 were sacrificed by day 42 and 54 respectively due to excessive tumor burden (>l,650 mm 3 ). In the group treated with four- arm 40K PEG-Gly-(7-ethyl-10-hydroxycamptothecin), only 1 of 7 animals was sacrificed on day 63.
• mice treated with four-arm 40K PEG-Gly-(7-ethyl-10- hydroxycamptothecin) had tumors ⁇ 1 ,650 mm 3 by day 72.
• the results show that four-arm 40K PEG-Gly-(7-ethyl-l 0-hydroxycamptotherin) has therapeutic activity in the treatment of CPT-11 refractory cancer.
• EXAMPLE 2 Therapeutic Efficacy of four-arm PEG-GIy-(7-ethyl-10-hydroxy- camptothecin) in Human Colorectal Tumor Xenografted Mice Refractory to CPT-H in the Second Round Treatment
• mice Human HT-29 colorectal tumors were established in nude mice by subcutaneous injection of 1 x 10 6 cells/mouse into a right auxiliary flank. When tumors reached an average volume of 100 mm 3 , mice were treated with CPT-1 1 (40 mg/kg/dose; q2d x 4). Mice were monitored for tumor growth. On day 15, mice that responded to CPT-11 therapy (mice that had tumor volumes ⁇ 3 x initial tumor volume at the start of CPT-11 therapy, i.e., mice considered CPT-sensitive) were selected, randomized and divided into two groups.
• One group was further treated with MTD of CPT-I I (40mg/kg/dose; q2d x 5) and the other group was treated with MTD of four-arm 40K PEG-Gly-(7-ethyl-10-hydroxycamptothecin) (10 mg/kg/dose q2d x 5) starting day 16.
• the drugs were administered intravenously via the tail vein.
• mice treated with CPT-11 tumor volume increased by 1298% compared to day 1.
• Mice treated with four-arm 40K PEG-Gly-(7- ethyl- 10-hydroxycamptothecin) had tumor volume moderately increased by 193%.
• 60% animals were sacrificed in CPT-11 treated group due to excessive tumor burden. No deaths have been recorded in four-arm 40IC PEG-Gly-(7-ethyl-10- hydroxycamptothecin) (compound 9) treated group on day 61.
• CEM/C2 CPT-refractory cell line
• CEM non-refractory parent cell line
• the CEM/C2 and CEM were obtained from NCI.
• the CEM cell lines are acute lymphoblastic leukemia cell lines. The in vitro cytotoxicity of each drug was determined using a MTS assay.
• cells were placed in 96-well plates (8 x 10 4 per well) and then treated with serial dilutions of four-arm 40IC PEG-Gly-(7-ethyl-10- hydroxycamptothecin) (compound 9), CPT or free 7-ethyl-lO-hydroxycamptothecin for 2 days at 37 0 C.
• MTS dye was added and incubated for 2 to 3 hours at 37 0 C and formation of a colored product (formazan) was measured at 490 nm.
• the % viability at each drug concentration was calculated as [OD test samples - background]/[OD controls (no treatment) - background].
• Sigmoidal dose response curves were generated by plotting Log (Drug) as a function of % viability (survival) and IC 50 values were calculated using the GraphPad Prism software. The results are set forth in Figures 3 and 4. The cytotoxicity ( ⁇ M of each compound that results in an IC 5 0) shows the in vitro anti-tumor potency of each compound. This study was used to determine the therapeutic effect of four-arm PEG-Gly ⁇ (7-ethyl-10- hydroxycamptothecin) in CPT-refractory cancer.
• the four-arm P EG-GIy- (7-efhyl- 10- hydroxy-camptothecin) was about 10 fold more potent than CPT-11 in the acute lymphoblastic leukemia cell line refractory to CPT as shown in Figure 3.
• all four-arm PEG-Gly-(7-ethyl-10-hydroxycamptothecin), CPT-I l and 7-ethyl-10- hydroxycamptothecin showed similar potency in the parent cell line (CEM) as shown in Figure 4.
• CEM parent cell line

Landscapes

• Health & Medical Sciences (AREA)
• Engineering & Computer Science (AREA)
• Bioinformatics & Cheminformatics (AREA)
• Life Sciences & Earth Sciences (AREA)
• General Health & Medical Sciences (AREA)
• Chemical & Material Sciences (AREA)
• Veterinary Medicine (AREA)
• Pharmacology & Pharmacy (AREA)
• Public Health (AREA)
• Animal Behavior & Ethology (AREA)
• Medicinal Chemistry (AREA)
• General Chemical & Material Sciences (AREA)
• Organic Chemistry (AREA)
• Chemical Kinetics & Catalysis (AREA)
• Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
• Epidemiology (AREA)
• Diabetes (AREA)
• Endocrinology (AREA)
• Oncology (AREA)
• Hematology (AREA)
• Reproductive Health (AREA)
• Emergency Medicine (AREA)
• Obesity (AREA)
• Pulmonology (AREA)
• Gynecology & Obstetrics (AREA)
• Pregnancy & Childbirth (AREA)
• Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
• Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
• Polyethers (AREA)

Priority Applications (9)

Applications Claiming Priority (2)

Publications (2)

Family

ID=39682430

Family Applications (1)

Country Status (13)

Cited By (4)

Families Citing this family (11)

Family Cites Families (59)

• 2008
  • 2008-02-08 EP EP08729405.4A patent/EP2109448B1/en not_active Not-in-force
  • 2008-02-08 WO PCT/US2008/053438 patent/WO2008098178A2/en not_active Ceased
  • 2008-02-08 CN CN2008800046084A patent/CN101605539B/zh not_active Expired - Fee Related
  • 2008-02-08 KR KR1020097016812A patent/KR20090108082A/ko not_active Ceased
  • 2008-02-08 RU RU2009133793/15A patent/RU2009133793A/ru unknown
  • 2008-02-08 AU AU2008213576A patent/AU2008213576B2/en not_active Ceased
  • 2008-02-08 MX MX2009008549A patent/MX2009008549A/es unknown
  • 2008-02-08 BR BRPI0807232-9A patent/BRPI0807232A2/pt not_active IP Right Cessation
  • 2008-02-08 JP JP2009549259A patent/JP2010518120A/ja active Pending
  • 2008-02-08 US US12/028,378 patent/US7928095B2/en not_active Expired - Fee Related
  • 2008-02-08 CA CA002677798A patent/CA2677798A1/en not_active Abandoned
  • 2008-02-12 TW TW097104779A patent/TW200902066A/zh unknown
• 2009
  • 2009-07-28 IL IL200100A patent/IL200100A0/en unknown
• 2011
  • 2011-04-18 US US13/088,825 patent/US20110195990A1/en not_active Abandoned

Non-Patent Citations (1)

Also Published As

Similar Documents

Legal Events