CN1391484A

CN1391484A - Method of potentiating chemotherapy and treating solid tumors

Info

Publication number: CN1391484A
Application number: CN00812921A
Authority: CN
Inventors: J·J·小吉邦斯; G·杜卡特; J·卢卡斯; L·A·斯派克
Original assignee: Wyeth LLC
Current assignee: Wyeth LLC
Priority date: 1999-09-15
Filing date: 2000-09-12
Publication date: 2003-01-15
Also published as: WO2001019399A3; EP1214092A2; JP2003509383A; AU7373600A; CA2381078A1; AR025659A1; BR0014001A; WO2001019399A2

Abstract

This invention provides a method of treating solid tumors which comprises administering an effective amount of a combination of (1) a bioresponse modifier and (2) a chemotherapeutic agent. This invention also provides a method of potentiating the effects of a chemotherapeutic regimen in a mammal in need of treatment with such regimen which comprises administering a bioresponse modifier in addition to a chemotherapeutic regimen.

Description

Methods for enhancing chemotherapeutic efficacy and treating solid tumors

The invention relates to the use of a combination of a biological response modifier and a chemotherapeutic agent for potentiation of chemotherapy and for the treatment of solid tumors.

Classical approaches to cancer chemotherapy for solid tumors have focused on the use of cytotoxic drugs, either targeted at essential metabolic processes (antimetabolites) or general toxins that interfere with multiple metabolic processes (alkylating agents). Antibiotic drugs (doxorubicin, mitoxantrone, mitomycin C, etc.) target the replication or integration of nucleic acids and are therefore, in general, toxic to cells. Similarly, microtubule active compounds (taxanes, vinblastines) also exhibit toxicity to cells, directed against their basic structural components.

The trend in recent years has been to combine drugs with different mechanisms of action, which has improved the biological response and survival rate of some solid tumors, including breast, prostate and small cell lung cancers. However, most solid tumors show low biological response rates and are not clearly visible to survival. Such cancers include non-small cell lung cancer, head and neck cancer, gastric cancer, pancreatic cancer, cervical cancer, melanoma, adrenocortical tumor, soft tissue sarcoma, and the like. In particular, new treatments are needed for these solid tumors and for solid tumors in general.

U.S. Pat. No. 5,312,831 (and EP652228) disclose carbamate and urea based drugs that induce cytokine production, for restoring neutrophils after cancer chemotherapy, radiation therapy, bone marrow transplantation or infection, for treating cancer, AIDS, aplastic anemia, myelodysplastic syndrome, infectious diseases, and for enhancing immune response.

U.S. Pat. No. 4,666,890 discloses a synthetic tripeptide that has been reported to act as an immunomodulator, as an antitumor agent rather than as a chemotherapeutic adjuvant. The reported cell wall components and their synthetic analogues are peptides incorporating the D-glutamic acid (D-Glu) moiety q-linked to lysine (Lys) or diaminopimelic acid (A2pm) together with other main peptide chains or fatty acyl groups flanking both ends.

Disclosure of Invention

The present invention provides a method of treating a solid tumor comprising combining effective amounts of (1) a biological response modifier and (2) a chemotherapeutic agent. The invention also provides methods of potentiating a chemotherapeutic regimen in a mammal in need of such treatment, which method comprises administering a biological response modifier in addition to the chemotherapeutic regimen. As used herein, the terms biological response modifier and chemotherapeutic agent include administration with one or more biological response modifiers and chemotherapeutic agents, e.g., the term chemotherapeutic agent can include administration with two chemotherapeutic agents.

Treatment is defined as providing a disease-modifying treatment or inhibiting the growth of, or effecting radical cure of, the solid tumor being treated.

For the purposes of the present invention within the definition of the scope of the invention, biological response modifiers are agents that activate the innate immune system in the body, typically including cytokine inducers and immune adjuvants. Cytokine inducers are substances that induce cytokine production, including cytokines such as IL-1, TNF; natural products such as muramyl dipeptide, lipopolysaccharides, and beta-glucan; synthetic cytokine inducers such as those disclosed in U.S. Pat. Nos. 5,312,831 and 4,666,890, the disclosures of which are incorporated herein by reference. Cytokine inducers are preferred biological response modifiers of the invention.

A particularly preferred cytokine inducing agent is of formula I disclosed in U.S. Pat. No. 5,312,831(EP652228) and has the structure:wherein,

R₁selected from hydrogen, substituted or unsubstituted (C)₁-C₂₀) An alkyl group, a substituted or unsubstituted cycloalkyl group, a substituted or unsubstituted cycloalkylalkyl group, a vinyl group, an ethynyl group, a substituted or unsubstituted amino group, a substituted or unsubstituted acylamino group, a substituted or unsubstituted aryl group, a substituted or unsubstituted aralkyl group, a substituted or unsubstituted aryloxy group, a substituted or unsubstituted alkoxyaryl group, a substituted or unsubstituted alkoxyaralkyl group, and a substituted or unsubstituted monocyclic or bicyclic heterocyclic group containing 1 to 4 hetero atoms selected from nitrogen, sulfur and oxygen atoms;

R_aand R₃Are respectively selected from hydrogen and substituted or unsubstituted (C)₁-C₆) Alkyl, substituted or unsubstituted alkoxyalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted cycloalkylalkyl, substituted or unsubstituted aryl, substituted or unsubstituted aralkyl, substituted or unsubstituted alkoxyaralkyl, vinyl, ethynyl, and substituted or unsubstituted monocyclic or bicyclic heterocyclic groups containing 1-4 heteroatoms selected from nitrogen, sulfur, and oxygen atoms, for R₃In which the heteroatom is not directly bonded to the-CH-group of the-CH-X-moiety;

R₂，R_band R_cRespectively selected from carboxyl or protected carboxyl, carboxyl or protected carboxyl lower alkyl and carboxyl amide;

x is oxygen or nitrogen;

R₄is H or an amino protecting group; wherein the substituents of the above-mentioned substituted alkyl, cycloalkyl, cycloalkylalkyl, amino, acylamino, aryl, aralkyl, aryloxy, alkoxyaryl, alkoxyaralkyl and heterocyclic groups are selected from the group consisting of halogen, hydroxy, lower alkyl, lower alkoxy, aryloxy, aralkoxy, amino, mono-or di-lower alkylamino, arylamino, aralkylamino, carboxy, formyl, lower alkoxycarbonyl, aryloxycarbonyl, aralkoxycarbonyl, lower alkylthio, arylthio, aralkylthio, arylsulfinyl, aralkylsulfinyl, lower alkylsulfonyl, arylsulfonyl, aralkylsulfonyl and monocyclic or bicyclic heterocyclic groups containing 1 to 4 hetero atoms selected from the group consisting of nitrogen, sulfur and oxygen;

or a pharmaceutically acceptable salt thereof.

The definition of the substituents and the preparation of the compounds of the formula I are described in U.S. Pat. No. 5,312,831 and EP 652228. The cytokine inducing agent of formula I may contain one or more asymmetric carbon atoms; in this case, the compounds of formula I include the various diastereomers, racemates and the various R and S isomers (entantiomers) thereof.

Various specific definitions of the above compounds and examples of these definitions are given below (taken from EP 652228B): a) (C)₁-C₂₀) The alkyl group may be a straight or branched lower alkyl group having 1 to 20 carbon atoms, such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl, neopentyl, isopentyl, hexyl, isohexyl, and the like. b) The cycloalkyl group may be a cycloalkyl group having 3 to 6 carbon atoms, such as cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl. c) The cycloalkylalkyl group may be a cycloalkylalkyl group having 4 to 12 carbon atoms, such as cyclopropylmethyl, cyclobutylmethyl, cyclopentylmethyl, cyclohexylmethyl, 1-cyclopropylethyl, 2-cyclopropylethyl1-cyclobutylethyl, 2-cyclobutylethyl, 1-cyclopentylethyl, 2-cyclopentylethyl, 1-cyclohexylethyl, 3-cyclohexylpropyl, 3-cyclopentylpropyl, 4-cyclohexylbutyl, 4-cyclopentylbutyl, 4-cyclopentylpentyl or 4-pentylcyclohexyl. d) The amido group can be an amido group in which the acyl moiety is derived from an acid, such as an organic carboxylic acid or carbonic acid. Each amido group includes an aliphatic group, an aryl group and/or a heterocyclic group in its molecule. These fatty acyl groups with acyl groups are derived from fatty acids and include: alkanoyl (e.g., formyl, acetyl, propionyl, butyryl, isobutyryl, valeryl, isovaleryl, pivaloyl, hexanoyl, α -ethyl-hexanoyl, heptanoyl, lauroyl, stearoyl, behenoyl), a group of formulae: CH (CH)₃(CH₂)₃₁CO，[CH₃(CH₂)₂₁]₂CHCO，[CH₃(CH₂)₁₅]₂CHCO，CH₃(CH₂)₄₁CO, etc.); lower alkoxycarbonyl (e.g., methoxycarbonyl, ethoxycarbonyl, propoxycarbonyl, butoxycarbonyl, t-pentyloxycarbonyl, etc.), etc. The acyl moiety may also be aroyl, the acyl group being derived from an acid of a substituted or unsubstituted aryl group, wherein the aryl group may include phenyl, tolyl, xylyl, naphthyl, and the like. Suitable examples thereof are as follows: aroyl (e.g., benzoyl, toluoyl, ditoluoyl, naphthoyl, phthaloyl, and the like); aralkoxycarbonyl (e.g., benzyloxycarbonyl, diphenylmethoxycarbonyl, triphenylmethoxycarbonyl, α -naphthyl-methoxycarbonyl, etc.) and the like. The acyl moiety may also be a heterocyclic acyl group, wherein the acyl group is derived from an acid having a heterocyclic ring, including: heterocyclic carbonyl, wherein the heterocyclic moiety is a 5-6 membered heterocyclic ring containing at least one heteroatom selected from the group consisting of nitrogen, oxygen and sulfur (e.g., thiophenoyl, furoyl, pyrrolecarbonyl, 5-oxo-2-pyrrolidinecarbonyl, nicotinoyl, etc.), and the like. e) The aryl group may be an aryl group having 6 to 15 carbon atoms, such as phenyl, biphenyl, 1-naphthyl or 2-naphthyl. f) The aralkyl group may be an aralkyl group having 7 to 15 carbon atoms, such as benzyl, 1-naphthylmethyl, 2-naphthylmethyl, 5, 6, 7, 8-tetrahydro-1-naphthyl, 5,6, 7, 8-tetrahydro-2-naphthyl, phenethyl, 3-phenylpropyl or 4-phenylbutyl. g) The aryloxy group may be an aryloxy group having 6 to 15 carbon atoms, such as a phenoxy group, a biphenyloxy group, a 1-naphthyloxy group, or a 2-naphthyloxy group. h) The alkoxyaryl or alkoxyaralkyl group may be an alkoxyaryl or alkoxyaralkyl group having 6 to 21 carbon atoms, such as benzoyl group, or alkoxybenzyl group. i) A monocyclic or bicyclic heterocyclic group containing 1 to 4 hetero atoms selected from nitrogen, sulfur and oxygen atoms, and may be a heterocyclic group having 4 to 15 carbon atoms such as pyrrolyl, furyl, thienyl, pyridyl, pyrimidyl, pyrazolyl, thiazolyl, isothiazolyl, isoxazolyl, oxazolyl, pyrazinyl, pyrimidinyl, pyridazinyl, indolyl, quinolyl, isoquinolyl, 2, 3-naphthyridinyl, quinoxalyl, quinazolinyl, 1, 4-benzodioxanyl, 1, 3-benzodioxanyl, 1, 2, 3-triazolyl, 1, 3, 4-thiadiazolyl, 1, 2, 3-thiadiazolyl, tetrazolyl, tetrahydrofuryl, tetrahydrothienyl, pyrrolidinyl, imidazolidinyl, 2-imidazolyl, etc, Morpholino, piperazinone N-oxide, piperazine N-oxide, morpholine N-oxide, lower alkyl morpholino such as N-methyl morpholino, N-ethyl morpholino or N-propyl morpholino, piperazinyl, piperidino, piperidinyl, thiomorpholino or thiomorpholino.

The aforementioned substituents (a) to (i) may be a halogen atom such as a chlorine atom, a fluorine atom or a bromine atom, a hydroxyl group; lower alkyl such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl or tert-butyl; lower alkoxy, such as methoxy, ethoxy, propoxy, isopropoxy, butoxy, isobutoxy, sec-butoxy or tert-butoxy; aryloxy such as phenoxy, 1-naphthoxy or 2-naphthoxy; aralkyloxy, such as benzyloxy, phenethyloxy, 1-naphthylmethoxy or 2-naphthylmethoxy; an amino group; mono-or di-lower alkylamino, such as methylamino, ethylamino, propylamino, isopropylamino, butylamino, sec-butylamino, isobutylamino, tert-butylamino, dimethylamino or diethylamino; arylamino, such as phenylamino, 1-naphthylamino or 2-naphthylamino; aralkylamino, such as benzylamino, phenethylamino, 1-naphthylmethylamino or 2-naphthylmethylamino; a carboxyl group; a formyl group; lower alkoxycarbonyl such as methoxycarbonyl, ethoxycarbonyl, propoxycarbonyl, isopropoxycarbonyl, butoxycarbonyl, sec-butoxycarbonyl, isobutoxycarbonyl or tert-butoxycarbonyl; an aryloxycarbonyl group such as a phenoxycarbonyl group, a 1-naphthyloxycarbonyl group or a 2-naphthyloxycarbonyl group; aralkoxycarbonyl, such as benzyloxycarbonyl, phenethyloxycarbonyl, 1-naphthylmethoxycarbonyl or 2-naphthylmethoxycarbonyl; a mercapto group; lower alkylthio such as methylthio, ethylthio, propylthio, isopropylthio, butylthio, sec-butylthio, isobutylthio or tert-butylthio; arylthio groups such as phenylthio, 1-naphthylthio or 2-naphthylthio; aralkylthio such as benzylthio, phenethylthio, 1-naphthylmethylthio or 2-naphthylmethylthio; an arylsulfinyl group such as a phenylsulfinyl group, a 1-naphthylsulfinyl group or a 2-naphthylmethionyl group; an aralkylsulfinyl group such as a benzylsulfinyl group, a phenethylsulfinyl group, a 1-naphthylmethylsulfinyl group or a 2-naphthylmethylsulfinyl group; lower alkylsulfonyl such as methylsulfonyl, ethylsulfonyl, propylsulfonyl, isopropylsulfonyl, butylsulfonyl, isobutylsulfonyl, sec-butylsulfonyl or tert-butylsulfonyl; arylsulfonyl such as phenylsulfonyl, 1-naphthalenesulfonyl or 2-naphthalenesulfonyl; aralkylsulfonyl such as benzylsulfonyl, phenethylsulfonyl, 1-naphthylmethylsulfonyl or 2-naphthylmethylsulfonyl; or a monocyclic or bicyclic heterocycle having from 4 to 15 carbon atoms and from 1 to 4 heteroatoms selected from oxygen, nitrogen and sulfur, such as pyrrolyl, furyl, thienyl, pyridyl, pyrimidinyl, pyrazolyl, thiazolyl, isothiazolyl, isoxazolyl, oxazolyl, pyrazinyl, pyrimidinyl, pyridazinyl, indolyl, quinolinyl, isoquinolinyl, 2, 3-naphthyridinyl, naphthridinyl, quinoxalinyl, quinazolinyl, 1, 4-benzodioxanyl, 1, 3-benzodioxanyl, 1, 2, 3-triazolyl, 1, 3, 4-thiadiazolyl, 1, 2, 3-thiadiazolyl, tetrazolyl, tetrahydrofuryl, tetrahydrothienyl, pyrrolidinyl, imidazolidinyl, 2-imidazolyl, morpholinyl, morpholino, morpholine N-oxide, morpholinyl, N-oxide, N-substituted, N-carbonyl, Lower alkylmorpholinyl, such as N-methylmorpholino, and N-ethylmorpholino or N-propylmorpholino, piperazinyl, piperidino, piperidinyl, thiomorpholinyl or thiomorpholinyl.

Cyclolower alkyl as used herein means C₁-C₆An alkyl group.

Protecting groups for protected carboxy or protected carboxy lower alkyl groups include the commonly used carboxy protecting groups conventionally used in peptide and amino acid chemistry by those skilled in the art, such as t.green: protecting Groups mentioned in protection Groups in Organic Synthesis, J Wiley and Sons, 1981. They include silyl esters, aliphatic esters and aryl esters such as trimethylsilyl, t-butyldimethylsilyl, acetyl, benzoyl and the like.

Protecting groups for protected amino groups include those commonly used in peptide and amino acid chemistry by those skilled in the art, such as those mentioned in t.green, supra, pages 218-287. The choice of a suitable protecting group is a condition for removal of the protecting group, consistent with other structural features of the compound. Suitable protecting groups include acyl groups such as t-butoxycarbonyl or benzyloxycarbonyl and the like.

Preferred compounds of formula I with respect to the general description above are those wherein:

R₁selected from substituted or unsubstituted (C)₁-C₂₀) An alkyl group, a substituted or unsubstituted cycloalkyl group, a substituted or unsubstituted cycloalkylalkyl group, a substituted or unsubstituted aryl group, a substituted or unsubstituted aralkyl group, a substituted or unsubstituted aryloxy group, a substituted or unsubstituted alkoxyaryl group, and a substituted or unsubstituted alkoxyaralkyl group. Wherein the aforementioned aryl group is selected from a substituted or unsubstituted phenyl group;

R_aand R₃Are respectively selected from hydrogen and substituted or unsubstituted (C)₁-C₆) An alkyl group;

x is oxygen or nitrogen;

R₄is H or an amino protecting group.

Furthermore, the most preferred compounds of formula I in the present invention are those wherein the following groups are present

R₁Is selected from (C)₄-C₁₄) Alkyl, cycloalkyl, (C)₂-C₈) Alkyl-substituted cycloalkyl, phenyl, benzyl, (C)₄-C₈) Alkylphenyl and (C)₁-C₆) Alkyl or alkoxy-benzyl;

R_aand R₃Independently selected from hydrogen and (C)₁-C₆) An alkyl group;

R₂，R_band R_cIndependently selected from carboxy or protected carboxy, carboxy or protected carboxy lower alkyl and carboxyamide;

x is oxygen or nitrogen;

R₄is H or an amino protecting group.

Particularly preferred compounds of formula I are those in which the following groups are present:

R₁selected from n-hexyl, 4-n-pentylcyclohexyl;

R₁and R₃Independently selected from hydrogen or methyl;

R₂，R_band R_cIs a carboxyl group;

x is oxygen or nitrogen;

R₄is H.

Particular preference is given to those compounds of the formula I having the D-isothreonine configuration:

wherein R is₃Is methyl, R₁And R₂As defined above.

It is also particularly preferred that the diaminopimediylalanine moiety of the molecule has the following stereochemistryThose compounds:

wherein R is₁Is methyl, R_b，R_cAnd R₄As defined above.

In terms of stereochemistry, the following compounds of formula I are most preferred:

wherein R is₃And R₁Is methyl, X is oxygen, R_b，R_c，R₁，R₂，R₄As defined above.

Particularly preferred cytokine inducers are [ R- (R, R) ]]-N- [ (R) -6-carboxy-N²- [ [ 2-carboxy-1-methyl-2- [ (1-oxoheptyl) amino group]Ethoxy radical]Carbonyl radical]-L-lysyl]Alanine or a pharmaceutically acceptable salt thereof (compound of formula I), which is disclosed in example 28 of U.S. patent No. 5,312,831.

U.S. Pat. No. 5,312,831 discloses several standard pharmacological test procedures that enable one skilled in the art to assess whether a compound is a biological response modifier. The induction of IL-6, CSF and G-CSF production was evaluated as described in the experimental procedures in columns 16-17.

Preferred chemotherapeutic agents are microtubule agents or macrophage activators. Microtubule active compounds are compounds that destabilize microtubules, either by preventing polymerization of tubulin (such as vincristine) or by preventing depolymerization of tubulin (such as taxanes). The assembly and disassembly of microtubules is a dynamic process, influenced by tubulin binding proteins and protein phosphorylation (kinases). Microtubule active compounds include taxanes such as paclitaxel or docetaxel, vincristine, vinblastine, vinorelbine and the like. Macrophage activators are compounds that activate macrophage activity, such as doxorubicin, doxirubicin and similar anthracenediones and anthracyclines, cisplatin, carboplatin, mitomycin C, bleomycin, and the like. Preferred chemotherapeutic agents are paclitaxel or carboplatin, or a combination of both. The microtubule agent or macrophage activating agent of the present invention can either be obtained commercially or can be prepared using standard preparation methods in the published literature.

The potentiating effect of biological response modifiers on chemotherapeutic agents has been evaluated in an in vivo standard pharmacological test procedure in which a human non-small cell lung cancer (NSCLC) cell line is engrafted into the flank of immunodeficient nude mice. The experimental procedures used and the results obtained are briefly described below. [ R- (R, R)]-N- [ (R) -6-carboxy-N²- [ [ 2-carboxy-1-methyl-2- [ (1-oxoheptyl) amino group]Ethoxy radical]Carbonyl radical]-L-lysyl]Alanine is used as a representative biological response modifier and paclitaxel is used as a representative chemotherapeutic agent.

6-8 week old Balb-c nu/nu female mice were obtained from the Charles Rive laboratory and 8-10 week old mice were used for the experiments. Human non-small cell lung cancer (NSCLC) cell line H-157 was from the American Tissue Culture Collection (ATCC) (Bethesda, Md.). Cells were cultured in RPMI medium with 10% fetal bovine serum.

Mice were injected subcutaneously with 7.5X 10⁶H-157 NSCLC tumor cells. When the tumor grows to 80-150mg, the length (l) and width (w) of the tumor are measured with a vernier caliper. Using the formula l × w²The volume of the tumor was calculated. In mm³The gauge volume can be directly converted to the assumed unit density in mg. 7 days after injection, the mice were randomized (day 0 of injection) into 10 groups, and the average tumor weight was 100 mg. Treatment groups were injected with paclitaxel (30mg/kg) on day 0 and drug vehicle (vehicle) or [ R (R, R) ]on days 1 and 8]-N- [ (R) -6-carboxy-N²- [ [ 2-carboxy-1-methyl-2- [ (1-oxoheptyl) amino group]Ethoxy radical]Carbonyl radical]-L-lysyl]Alanine (100. mu.g/kg).

On days 7 and 14, paclitaxel plus [ R (R, R) ]was received]-N- [ (R) -6-carboxy-N²- [ [ 2-carboxy-1-methyl-2- [ (1-oxoheptyl) amino group]Ethoxy radical]Carbonyl radical]-L-lysyl]The growth rate of the tumors in the alanine group was significantly inhibited compared to the untreated control group or the group treated with paclitaxel alone. The inhibition of tumor growth by paclitaxel alone (p.ltoreq.0.05 by student's t) was measured on day 7, and [ R- (R, R).)]-N- [ (R) -6-carboxyradical-N²- [ [ 2-carboxy-1-methyl-2- [ (1-oxoheptyl) amino group]Ethoxy radical]Carbonyl radical]-L-lysyl]Alanine, the degree of inhibition is significantly enhanced. On day 14, paclitaxel alone showed no significant difference in inhibition from the control group; and receiving paclitaxel on day 0 and injecting on days 1 and 8 [ R- (R, R)]-N- [ (R) -6-carboxy-N²- [ [ 2-carboxy-1-methyl-2- [ (1-oxoheptyl) amino group]Ethoxy radical]Carbonyl radical]-L-lysyl]The group with-alanine still had significant inhibitory effect compared to the untreated control group or the group with paclitaxel alone. The results are shown in Table 1. In the following table, [ R (R, R).)]-N- [ (R) -6-carboxy-N²- [ [ 2-carboxy-1-methyl-2- [ (1-oxoheptyl) amino group]Ethoxy radical]Carbonyl radical]-L-lysyl]The code of alanine is CI.

TABLE 1 inhibition of tumor cell growth by CI plus paclitaxel in nude mice xenografts

Treatment group	^aTumor weight (mg) day 7	% inhibition	Tumor weight (mg) day 14	% inhibition
Treatment group	^aTumor weight (mg) day 7	% inhibition	Tumor weight (mg) day 14	% inhibition	Blank control group	410±99	-	1234±354	-
Paclitaxel group	^c289±108	30	866±348	30	Blank control group	410±99	-	1234±354	-
Paclitaxel group	^c289±108	30	866±348	30	Paclitaxel + CI group	^b182±69	56	^b491±151	60

^aData were from 5 separate evaluations (mean ± standard deviation), at each evaluation, 10 mice per group. Treatment groups received paclitaxel (30mg/kg) on day 0 and injected with solvent or CI (100 μ g/kg) on days 1 and 8.^bP.ltoreq.0.05 compared to the blank control or paclitaxel alone (Student's t test).^cP is less than or equal to 0.05 compared with a blank control group.

Only [ R- (R, R). ] on days 1 and 8]-N- [ (R) -6-carboxy-N²- [ [ 2-carboxy-1-methyl-2- [ (1-oxoheptyl) amino group]Ethoxy radical]Carbonyl radical]-L-lysyl]Evaluation of tumor-bearing mice with-alanine (100. mu.g/kg) was carried out several times (data not shown), none of which showed the use of [ R- (R, R).)]-N- [ (R) -6-carboxy-N²- [ [ 2-carboxy-1-methyl-2- [ (1-oxoheptyl) amino group]Ethoxy radical]Carbonyl radical]-L-lysyl]Alanine itself has an inhibitory effect on the growth of tumor cells in nude mice. [ R- (R, R)]-N- [ (R) -6-carboxy-N²- [ [ 2-carboxy-1-methyl-2- [ (1-oxoheptyl) amino group]Ethoxy radical]Carbonyl radical]-L-lysyl]Alanine does not directly inhibit tumor cell growth in vitro, nor does it enhance the inhibition of paclitaxel on tumor cell growth in cell culture.

Clinical studies have also been conducted to evaluate the effects of biological response modifiers and chemotherapeutic agents. [ R- (R, R)]-N- [ (R) -6-carboxy-N²- [ [ 2-carboxy-1-methyl-2- [ (1-oxoheptyl) amino group]Ethoxy radicalBase of]Carbonyl radical]-L-lysyl]Alanine as a representative biological response modifier, paclitaxel and carboplatin as representative chemotherapeutic agents. In this study, patients with advanced cancer were given [ R- (R, R) about 7 days prior to the first round of chemotherapy (carboplatin and paclitaxel)]-N- [ (R) -6-carboxy-N²- [ [ 2-carboxy-1-methyl-2- [ (1-oxoheptyl) amino group]Ethoxy radical]Carbonyl radical]-L-lysyl]Alanine (round 0). Patients were given a second round of chemotherapy about 21 days later, and given [ R- (R, R). ]on days 1 and 8 after the second round of chemotherapy]-N- [ (R) -6-carboxy-N²- [ [ 2-carboxy-1-methyl-2- [ (1-oxoheptyl) amino group]Ethoxy radical]Carbonyl radical]-L-lysyl]Alanine, at the doses indicated in the table below. 69% of patients are clinically effective, with tumors either disappearing completely, or partially diminishing, or stabilizing the disease. Of the 16 treated patients, 6 were proposed to be diagnosed with NSCLC. In this group, 3 responded completely, 1 responded partially, 1 was stable, and only 1 developed. The expected complete response rate of chemotherapy with standard chemotherapy regimens (paclitaxel and carboplatin) is only about 5%, and it is completely unexpected that 3 of 6 treated patients show a complete response. In this treatment regimen, the paclitaxel dose is 175-200mg/m²The area under the curve (AUC) dose of carboplatin was 6mg/ml × min. The results of the clinical trials are summarized in the table below. In the following table, [ R- (R, R).)]-N- [ (R) -6-carboxy-N²- [ [ 2-carboxy-1-methyl-2- [ (1-oxoheptyl) amino group]Ethoxy radical]Carbonyl radical]-L-lysyl]Alanine is denoted by CI.

TABLE 2 treatment results for patients with advanced cancer with combinations of paclitaxel, carboplatin and CI

			^aTumor response
			^aTumor response				# treated patient	Dosage (μ g/kg)^b	Is totally produced from	In part	Stabilization	Development of
							# treated patient	Dosage (μ g/kg)^b	Is totally produced from	In part	Stabilization	Development of
						3	0.1	2	1
6	0.2		1	2	3	3	0.1	2	1
6	0.2		1	2	3	4	0.266	1	1	1	1
3	0.4	1		1	1	4	0.266	1	1	1	1
3	0.4	1		1	1
Total up to	16		4	3	4							5

^aCompletely: without obvious symptoms

The method comprises the following steps: the tumor reduction is more than or equal to 50 percent

And (3) stabilizing: the disease condition is not further developed

And (3) development: the disease condition is developed

^bCI dose

The results of in vivo standard pharmacological and clinical trials with representative biological response modifiers and chemotherapeutic agents indicate that the combination of biological response modifiers with chemotherapeutic agents can be used to enhance the efficacy of standard chemotherapy for the treatment of solid tumors. In particular, the compositions of the present invention are useful for the treatment of non-small cell lung cancer, glioma, ovarian cancer, breast cancer, prostate cancer, head and neck cancer, renal cancer, pancreatic cancer, liver cancer, colon cancer, soft tissue sarcoma and the like.

The combination therapies used in the present invention may be administered simultaneously, or may be staggered. The biological response modifier may be administered at different times during the course of chemotherapy. Thus, the term combination does not imply that the agents must be administered simultaneously, or as a whole. A typical useful treatment regimen, for example in clinical studies, is that a patient receives a biological response modifier therapy prior to receiving a first course of chemotherapy. Typically, the chemotherapy is repeated several weeks thereafter, in which case the biological response modifier may be administered one or more days after the second course of chemotherapy. This staggered administration may continue throughout the treatment period.

For simultaneous administration, the combined ingredients may be formulated in a single dosage form, but the combined ingredients are all pre-formulated for separate administration.

Formulations of specialized chemotherapeutic agents are well known in the art (and most are commercially available or are formulated for use in clinical trials), and these agents can be administered typically intravenously or orally; however, depending on each agent and patient, the chemotherapeutic agent may also be administered parenterally, rectally, vaginally, transdermally, subcutaneously, topically, nasally, or by direct infusion at the site of the cancerous lesion.

The biological response regulator of the invention can also be taken orally, intravenously, parenterally, rectally, vaginally, transdermally, subcutaneously, topically, nasally or by direct infusion at the site of cancerous lesions. Biological response modifiers can be formulated according to standard literature methodologies. For example, compounds of formula I and [ R- (R, R)]-N- [ (R) -6-carboxy-N²- [ [ 2-carboxy-1-methyl-2- [ (1-oxoheptyl) amino group]Ethoxy radical]Carbonyl radical]-L-lysyl]The formulation of alanine is described in U.S. Pat. No. 5,312,831.

Biological response modifiers that are not commercially available or available in clinical trials may be formulated alone with the chemotherapeutic agent (or resulting unsuitable for use) or in combination with one or more pharmaceutically acceptable carriers for administration, e.g., solvents, diluents, and the like. Can be administered orally in the form of tablet, capsule, dispersible powder, granule, suspension, etc., such as suspension in an amount of 0.05% -5%, syrup containing 10% -50% sugar, elixir containing 20% -50% ethanol, etc., or parenteral sterile injection solution or suspension in isotonic medium containing 0.05% -5%. Such pharmaceutical formulations may contain from 0.05% to 90% of the active ingredient in combination with a carrier. Usually, the content is at most 5 to 60% by weight.

Solid carriers which may be present in the formulations for tablet and capsule administration include starch, lactose, dicalcium phosphate, microcrystalline cellulose, sucrose and kaolin, and liquid carriers may include sterile water, polyethylene glycols, nonionic surfactants and edible oils such as corn, peanut and sesame oils, as are appropriate to the nature of the active ingredient and the particular dosage form for which administration is desired. Adjuvants commonly used in the preparation of pharmaceutical compositions may be flavouring agents, colouring agents, preservatives and antioxidants, such as vitamin E, vitamin C, BHT and BHA.

The components of the combination may be administered by injection or intraperitoneally. These active ingredients may be prepared as solutions or suspensions in water in the form of the free base or pharmaceutically acceptable salts, mixed with a suitable surfactant, such as hydroxypropylcellulose. The dispersant may be prepared from glycerol, liquid polyethylene glycol or a mixture of both in oil. Under normal conditions of storage and use, these preparations contain a preservative to prevent the growth of microorganisms.

Pharmaceutical dosage forms suitable for injectable use include sterile aqueous solutions or dispersions and sterile powders for the extemporaneous preparation of sterile solutions or dispersions. Each injectable form must be sterile and must have a certain fluidity so that it can be easily placed in a syringe. The formulations must be stable under the conditions of manufacture and storage and must be resistant to contamination by microorganisms, such as bacteria, molds, and the like. The carrier must be a solvent or dispersion medium such as water, alcohol, polyol (e.g., glycerol, propylene glycol, and liquid polyethylene glycol), suitable mixtures thereof, and vegetable oils.

It is contemplated that the dosages of chemotherapeutic agents and biological response modifiers used during treatment will be adjusted based on the patient's inherent toxic response to treatment and to standard chemotherapeutic methods. Such as by reducing the amount of chemotherapeutic agent, depending on myelosuppression or decreased liver function. The starting dose of chemotherapeutic agent is expected to be the starting amount without the biological response modifier present. For example, for paclitaxel, carboplatin and [ R- (R, R)]-N- [ (R) -6-carboxy-N' - [ [ 2-carboxy- ]1-methyl-2- [ (1-oxoheptyl) amino group]Ethoxy radical]Carbonyl radical]-L-lysyl]-alanine combination therapy, with estimated paclitaxel amount of 135-225mg/m²The preferred dosage is 175-200mg/m²The area under the curve (AUC) dose of carboplatin is 5-7mg/ml × min, with a preferred amount of 6mg/ml × min. The designed dose of the biological response modifier varies with its strength to induce the immune system. In the case of cytokine inducers, they vary according to their ability to induce cytokine production. For [ R- (R, R)]-N- [ (R) -6-carboxy-N²- [ [ 2-carboxy-1-methyl-2- [ (1-oxoheptyl) amino group]Ethoxy radical]Carbonyl radical]-L-lysyl]Alanine, the initial dose expected to range from 0.005 to 2. mu.g/kg, the preferred dose being from 0.01 to 1. mu.g/kg.

Claims

1. A method of treating a solid tumor in a mammal, comprising administering to said mammal an effective amount of a combination of a biological response modifier and a chemotherapeutic agent.

2. Use of a biological response modifier and a chemotherapeutic agent in the preparation of a medicament for the treatment of solid tumors in a mammal.

3. The method or use of claim 1 or 2, wherein the biological response modifier is a cytokine inducer.

4. The method or use of claim 3, wherein the cytokine inducer is a compound of formula I, having the structure:

wherein,

R_aand R₃Independently selected from hydrogen, substituted or unsubstituted (C)₁-C₆) Alkyl, substituted or unsubstituted alkoxyalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted cycloalkylalkyl, substituted or unsubstituted aryl, substituted or unsubstituted aralkyl, substituted or unsubstituted alkoxyaralkyl, vinyl, ethynyl, and substituted or unsubstituted monocyclic or bicyclic heterocyclic groups containing 1-4 heteroatoms selected from nitrogen, sulfur, and oxygen atoms, for R₃In which the heteroatom is not directly bonded to the-CH-group of the-CH-X-moiety;

x is oxygen or nitrogen;

R₄is H or an amino protecting group; wherein the substituent for the above-mentioned substituted alkyl, cycloalkyl, cycloalkylalkyl, amino, acylamino, aryl, aralkyl, aryloxy, alkoxyaryl, alkoxyaralkyl and heterocyclic group is selected from the group consisting of halogen, hydroxy, lower alkyl, lower alkoxy, aryloxy, aralkyloxy, amino, mono-or di-lower alkylamino, arylamino, aralkylamino, carboxy, formylLower alkoxycarbonyl, aryloxycarbonyl, aralkoxycarbonyl, lower alkylthio, arylthio, aralkylthio, arylsulfinyl, aralkylsulfinyl, lower alkylsulfonyl, arylsulfonyl, aralkylsulfonyl, and monocyclic or bicyclic heterocyclic group containing 1 to 4 heteroatoms selected from nitrogen, sulfur and oxygen; or a pharmaceutically acceptable salt thereof.

5. The method or use of claim 4, wherein the compound of formula I is [ R- (R, R) ]]-N- [ (R) -6-carboxy-N²- [ [ 2-carboxy-1-methyl-2- [ (1-oxoheptyl) amino group]Ethoxy radical]Carbonyl radical]-L-lysyl]-alanine or a pharmaceutically acceptable salt thereof.

6. The method or use of any one of claims 1-5, wherein the chemotherapeutic agent is a microtubule agent or a macrophage activator.

7. The method or use of claim 6 wherein the microtubule agent or macrophage activating agent is selected from the group consisting of paclitaxel, docetaxel, vincristine, vinblastine, vinorelbine, doxorubicin, doxirubicin, cisplatin, carboplatin, mitomycin C and bleomycin.

8. The method or use of claim 7, wherein the microtubule agent or macrophage activating agent is paclitaxel and carboplatin.

9. A method of potentiating chemotherapy in a mammal in need of chemotherapy, which method comprises administering a biological response modifier in addition to chemotherapy.

10. The method of claim 9, wherein the biological response modifier is a cytokine inducer.

11. The method of claim 10, wherein the cytokine inducing agent is a compound of formula I having the structure:wherein,

x is oxygen or nitrogen;

R₄is H or an amino protecting group; wherein the substituent for said substituted alkyl, cycloalkyl, cycloalkylalkyl, amino, acylamino, aryl, aralkyl, aryloxy, alkoxyaryl, alkoxyaralkyl and heterocyclic group is selected from the group consisting of halogen, hydroxy, lower alkyl, lower alkoxy, aryloxy, aralkyloxy, amino, mono-or di-lower alkylamino, arylamino, aralkylamino, carboxy, formyl, lower alkoxycarbonyl, aryloxycarbonyl, aralkoxycarbonyl, lower alkylthio, arylthio, aralkylthio, arylsulfinyl, aralkylsulfinyl, lower alkylsulfonyl, arylsulfonyl, aralkylsulfinyl, alkoxyaryl, alkoxyaralkyl and heterocyclic groupArylsulfonyl, aralkylsulfonyl and monocyclic or bicyclic heterocyclic groups containing 1 to 4 heteroatoms selected from nitrogen, sulfur and oxygen; or a pharmaceutically acceptable salt thereof.

12. The method of claim 11, wherein the compound of formula I is [ R- (R, R) ]]-N- [ (R) -6-carboxy-N²- [ [ 2-carboxy-1-methyl-2- [ (1-oxo-heptyl) amino group]Ethoxy radical]Carbonyl radical]-L-lysyl]-alanine or a pharmaceutically acceptable salt thereof.

13. The method of any one of claims 9-12, wherein the chemotherapeutic agent is a microtubule agent or a macrophage activator.

14. The method of claim 13, wherein the microtubule agent or macrophage activating agent is selected from the group consisting of paclitaxel, docetaxel, vincristine, vinblastine, vinorelbine, doxorubicin, doxirubicin, cisplatin, carboplatin, mitomycin C, and bleomycin.

15. The method of claim 14, wherein the microtubule agent or macrophage activating agent is paclitaxel and carboplatin.

16. Use of a product containing a biological response modifier and a chemotherapeutic agent as a combined preparation for simultaneous, separate or sequential administration in the treatment of a solid tumor.

17. The product of claim 16, wherein the biological response modifier is a cytokine inducer.

18. The article of manufacture of claim 17, wherein the cytokine inducer is a compound of formula I having the structure:

wherein,

R₁selected from hydrogen, substituted or unsubstituted (C)₁-C₂₀) Alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted ringAn alkyl group, a vinyl group, an ethynyl group, a substituted or unsubstituted amino group, a substituted or unsubstituted acylamino group, a substituted or unsubstituted aryl group, a substituted or unsubstituted aralkyl group, a substituted or unsubstituted aryloxy group, a substituted or unsubstituted alkoxyaryl group, a substituted or unsubstituted alkoxyaralkyl group, and a substituted or unsubstituted monocyclic or bicyclic heterocyclic group containing 1 to 4 hetero atoms selected from nitrogen, sulfur and oxygen atoms;

x is oxygen or nitrogen;

R₄is H or an amino protecting group; wherein the substituents of the above-mentioned substituted alkyl, cycloalkyl, cycloalkylalkyl, amino, acylamino, aryl, aralkyl, aryloxy, alkoxyaryl, alkoxyaralkyl and heterocyclic groups are selected from the group consisting of halogen, hydroxy, lower alkyl, lower alkoxy, aryloxy, aralkoxy, amino, mono-or di-lower alkylamino, arylamino, aralkylamino, carboxy, formyl, lower alkoxycarbonyl, aryloxycarbonyl, aralkoxycarbonyl, lower alkylthio, arylthio, aralkylthio, arylsulfinyl, aralkylsulfinyl, lower alkylsulfonyl, arylsulfonyl, aralkylsulfonyl and monocyclic or bicyclic heterocyclic groups containing 1 to 4 hetero atoms selected from the group consisting of nitrogen, sulfur and oxygen; or a pharmaceutically acceptable salt thereof.

19. The process of claim 18The product of (1), wherein the compound of formula I is [ R- (R, R')]-N- [ (R) -6-carboxy-N²- [ [ 2-carboxy-1-methyl-2- [ (1-oxoheptyl) amino group]Ethoxy radical]Carbonyl radical]-L-lysyl]-alanine or a pharmaceutically acceptable salt thereof.

20. The product of claims 16-19, wherein the chemotherapeutic agent is a microtubule agent or a macrophage activator.

21. A product as claimed in claim 20 wherein the microtubular agent or macrophage activating agent is selected from paclitaxel, docetaxel, vincristine, vinblastine, vinorelbine, doxorubicin, doxirubicin, cisplatin, carboplatin, mitomycin C and bleomycin.

22. The product of claim 20, wherein the microtubular agent or macrophage activating agent is paclitaxel and carboplatin.