CZ20003037A3 - Antitumor substances - Google Patents

Abstract

Antitumor agents of formula (I) or (II) wherein the individual general symbols have the meaning given to these of the formulas are compounds with a very good effect against cancers including solid tumors and tumors, resistant to commonly used chemotherapeutic agents. It also forms part of the solution a pharmaceutical composition comprising said compounds.

Description

Technical field

The invention relates to a class of antitumor agents and pharmaceutical compositions containing them.

BACKGROUND OF THE INVENTION

Some malignant tumors, in which chemotherapy is used predominantly to prolong life expectancy, have already achieved considerable success. These include Burkitt's lymphoma, acute lymphocytic leukemia and Hodgkin's sarcoma, and 10 to 15 other tumor types. These tumors have been described, for example, in A. Golden et al., Eur. J. Cpncer, 17, 129, 1981 in Table 1. However, these tumors, in which the patient's lifespan and quality of life have been substantially prolonged, represent only a small fraction of the various types of cancer. Furthermore, there are very few substances that are truly highly effective against solid tumors, such as ovarian, mammary, lung, etc. Cyclophosphamide, adriamycin, 5-FU, hexamethylmelamine and the like. there is a significant risk of relapse and increased mortality in patients with many types of cancer.

In the event of relapse, remission may be achieved in some patients by other treatments. However, in this case, it is usually necessary to use high doses of the original chemotherapeutic agent or additionally to use additional agents since the tumor develops at least partial resistance to the agents used. Recent research has shown that resistance can develop simultaneously against several active substances, including those that have not yet been used in a patient. The formation of resistant tumors may be a function of tumor mass • ·

and is the main cause of unsuccessful healing. In order to overcome this resistance, chemitherapy at high doses or doses can be used. along with irradiation and allogeneic or autologous bone marrow transplantation. When using high doses of chemicals, the parent compound may be used or other substances may be administered. Obviously, new agents that do not cross resistance to mdr phenotypes should be discovered in the development of new agents, otherwise treatment success and easier treatment in previously treated patients cannot be improved.

The antitumor activity of a new class of natural substances, called illudins, was investigated in vitro by Kelner M. et al., Cancer Res., 47, 3186, 1987. Illudin M was purified and tested for in vivo efficacy at the National Cancer Institute Division of Cancer Treatment NCI DCT. Illudine M has been shown to significantly increase life expectancy in Dunning leukemia rats, but has a significantly lower therapeutic index for solid tumors. The high toxicity of these substances did not allow any human testing. Recently, synthetic illudine analogs have been developed with promising antitumor activity, including those described in US 5439936 and 5523490.

Clearly, despite considerable progress, there is still a need to find new chemotherapeutic agents capable of inhibiting tumor growth, particularly in solid tumors. At the same time, the new agents should have a therapeutic index that can be used in in vivo treatment.

SUMMARY OF THE INVENTION

The present invention provides anticancer compounds of the formula I.

where

R 1 represents a hydrogen atom, a hydroxy group, a mercapto group, an amino group, a halogen atom, a carboxy group, a nitro group or a - (CH ₂ ) n - (X) - (Y) group.

n is an integer from 0 to 4,

X is -O-, thio -S-, -N (R _a ) - or absent,

Y is C 3 -C 6 cycloalkyl, aryl, heteroaryl, a saccharide, amino acid, peptide residue or a 1 to 15-membered branched or unbranched hydrocarbon chain, optionally containing 1, 2 or 3 non-peroxide oxy, thio or -N (R _a ) - groups, wherein: this chain is optionally substituted on a carbon atom with 1, 2 or 3 substituents from oxo = O, hydroxy, carboxy, halogen, mercapto, nitro, -N (R b) _c , C 3 -C 6 cycloalkyl, aryl, heteroaryl, saccharide residue, amino acids or peptides and at the same time the chain is saturated or unsaturated,

R ₂ is carboxy, C 1 -C 6 alkanoyl, C 1 -C 6 alkoxycarbonyl, halo C1-C6 alkyl, -C (= O) NR d R e, a saccharide residue, amino acids, peptides, or C1-C6 alkyl substituted by 1 or 2 hydroxy, C1-C6alkoxy,

C1-C6alkanoyloxy, carboxy, amino acid, peptide or carbohydrate residues or -C (= O) NRdR _e >

R ₃ is hydrogen, C 1 -C 6 alkyl, C 1 -C 6 alkoxy,

C 1 -C 6 alkylthio, aryl, heteroaryl, aryloxy or heteroaryloxy,

R ₄ is hydrogen or C 1 -C 6 alkyl, R ₅ is hydroxy, C 1 -C 6 alkoxy or C 1 -C 6 alkanoyloxy or R ₄ and R ₅ together form ethylenedioxy,

R ₆ is hydrogen, carboxy, C 1 -C 6 alkanoyl, C 1 -C 6 alkoxycarbonyl, halogenC 1 -C 6 alkyl, -C (= O) NR _f R _g , a saccharide, amino acid or peptide residue or C 1 -C 6 alkyl optionally substituted with 1 to 2 substituents from the group hydroxy, C1-C6alkoxy, C1-C6alkanoyloxyskupina, carboxy, amino acid residue, peptide or carbohydrate or -C (= O) NR _f R _g,

R _a represents a hydrogen atom, C 1 -C 6 alkyl, C 1 -C 6 alkanoyl, phenyl or benzyl a

R b, R c, Rd, R _e , R f 3 R _g independently represent hydrogen, C 1 -C 6 alkyl, C 1 -C 6 alkanoyl, phenyl or benzyl or form R b and R _c or R _d and R _e or R f and R _g together with the nitrogen atom to which are attached to form pyrrolidine, piperidine or morpholine ring wherein any aryl, heteroaryl, aryloxy or heteroaryloxy in the definition of Y or R ₃ is optionally substituted with 1, 2 or 3 substituents selected from C1-C6alkyl, C1-C6alkoxy, C1 6 alkanoyl, C1- C6alkanoyloxy, C1-C6alkoxycarbonyl, hydroxyC1-C6alkyl, halogenC1-C6alkyl, hydroxy, halogen, carboxy, mercapto, nitro or -N (Rh) (Rj), wherein Rh and Rj independently represent hydrogen, C1-C6alkyl, C1-C6alkanoyl , phenyl or benzyl, or together with the nitrogen atom to which they are attached a pyrrolidine, piperidine or morpholine ring, as well as pharmaceutically acceptable salts thereof.

The present invention also relates to those compounds of formula I wherein R 1 is - (CH 2) _n - (X) - (O, where n is

0 to 4, X stands for oxy, thio, -N (R _a ) - or absent, Y stands for amino acid residue protected by 1 or 2 protecting groups, peptide residue or one to 15-membered branched or unbranched hydrocarbon chain, optionally containing 1 , 2 or 3 non-peroxide oxy, thio or -N (R _a ) -, this chain is substituted with 1, 2 or 3 peptide residues and is saturated or unsaturated,

R ₂ is hydrogen or C1-C6 alkyl, _R3 is hydrogen, C1 -C6 alkyl, C1 6 alkoxy, C1 C6alkylthioskupinu, aryl, heteroaryl, aryloxy or heteroaryloxy; R4 is hydrogen or C1 -C6 alkyl and R5 is hydroxy, C1 -C 6 alkoxy or C 1 -C C6alkanoyloxyskupinu forms or R4 and _R5 together form ethylenedioxy, R g is hydrogen, carboxy, C 1 -C 6 alkanoyl,

C 1 -C 6 alkoxycarbonyl, halogenC 1 -C 6 alkyl, -C (= O) NR _f R _g , carbohydrate, amino acid or peptide residue or C 1 -C 6 alkyl optionally substituted with 1 to 2 substituents from hydroxy, C 1 -C 6 alkoxy, C 1 -C 6 alkanoyloxy, carboxy, amino acid , peptides or carbohydrate or the group -C (= O) NR _f R _g ,

R _a represents a hydrogen atom, C 1 -C 6 alkyl, C 1 -C 6 alkanoyl, phenyl or benzyl a

R b, R c, Rd, R e, R _f and R _g independently represent hydrogen, C 1 -C 6 alkyl, C 1 -C 6 alkanoyl, phenyl or benzyl or form R b and R _c or Rd and R _e or R f and R _g together with the nitrogen atom, to which are bound a pyrrolidine, piperidine or morpholine ring, wherein any aryl, heteroaryl, aryloxy or heteroaryloxy group Y or R ₃ is optionally substituted with 1, 2 or 3 substituents from C 1 -C 6 alkyl, C 1 -C 6 alkoxy, C 1 -C 6 alkanoyl, C1-C6alkanoyloxy, C1-C6alkoxycarbonyl, hydroxyC1-C6alkyl, halogenC1-C6alkyl, hydroxy, halogen, carboxy, mercapto,

nitro or -N (Rh) (R j) wherein R _h and R j independently represent hydrogen, C 1 -C 6 alkyl, C 1 -C 6 alkanoyl, phenyl or benzyl or form together with the nitrogen atom to which they are attached a pyrrolidine, piperidine or morpholine ring, as well as pharmaceutically acceptable salts thereof. A preferred value for Y is C 1 -C 6 alkyl, substituted with a peptide residue.

The present invention also provides a compound of formula I wherein R 1 is hydrogen, hydroxy, mercapto, amino, halogen, carboxy, nitro or - (CH ₂ ) _n - (X) - (Y), n is an integer of 0 to 4, X is oxy, thio, -N (R _a ) - or absent, Y is C 3 -C 6 cycloalkyl, aryl, heteroaryl, a saccharide, amino acid or peptide residue or a 1 to 15-membered branched or unbranched carbon chain, optionally containing 1 , 2 or 3 non-peroxide oxy, thio or -N (R _a ) - groups, the chain being optionally substituted by 1, 2 or 3 oxo, hydroxy, carboxy, halogen, mercapto, nitro,

-N (R _b ) (R _c ), C 3 -C 6 cycloalkyl, aryl, heteroaryl, a saccharide, amino acid or peptide residue, the chain may be saturated or unsaturated, R ₂ is hydrogen or C 1 -C 6 alkyl, R ₃ is hydrogen, C1 6 alkyl, C 1 -C 6 alkoxy, C 1 -C C6alkylthioskupinu, aryl, heteroaryl, aryloxy or heteroaryloxy; R ₄ is hydrogen or a C1-C6alkyl, R5 is hydroxy, C 1 -C 6 alkoxy or C 1 -C C6alkanoyloxyskupinu or _R4 and R5 together form ethylenedioxy, R6 is carboxy, C 1 -C 6 alkanoyl, C 1 -C 6 alkoxycarbonyl, -C (= O) NR _f R _g, a saccharide residue, amino acid or peptide or a C1-C6alkyl, substituted with 1 or 2 substituents selected from C1-C6alkoxy,

C 1 -C 6 alkanoyloxy, carboxy, amino acid, peptide or carbohydrate residue or -C (= O) NR _f R _g , R _a represents hydrogen, ·· ·· ► * * | · · ·· · ·

C 1 -C 6 alkyl, C 1 -C 6 alkanoyl, phenyl or benzyl and R _b , R _c , Rd, R e, R f and R g independently represent hydrogen, C 1 -C 6 alkyl, C 1 -C 6 alkanoyl, phenyl or benzyl or form R _b and R _c , or R _d and R _e or R f and R _g together with the nitrogen atom to which the pyrrolidine, piperidine or morpholine ring are bonded, wherein any aryl, heteroaryl, aryloxy or heteroaryloxy group of Y or R ₃ is optionally substituted with 1, 2 or 3 substituents C1-C6alkyl, C1-C6alkoxy, C1-C6alkanoyl,

C 1-6 alkanoyloxy, C 1 -C 6 alkoxycarbonyl, hydroxyC 1 -C 6 alkyl, halogenC 1 -C 6 alkyl, hydroxy, halogen, carboxy, mercapto, nitro or -N (R _h ) (R j), where R _h and R j independently represent hydrogen, C 1 -C 6 alkyl, C 1 -C 6 alkanoyl, phenyl or benzyl or together with the nitrogen atom to which they are attached a pyrrolidine, piperidine or morpholine ring, as well as pharmaceutically acceptable salts thereof.

The invention also relates to dimeric compounds consisting of two compounds of formula I linked by a linker. The group may be, for example, an alkyl type or an ester type. Examples include - (CH ₂ ) p O - (CH ₂ ) q -, - (CH ₂ ) _r - and -CH ₂ -S-CH ₂ C (O) -O- (CH ₂ ) ₂ -OC (O) CH ₂ -S-CH ₂ -, where p and q are integers from 1 to 8 and r is an integer from 1 to 16, preferably 1 to 8. It will be appreciated that other groups of similar length may be used. Two compounds of formula I can be combined e.g., by one of the symbols R, R _3, R 4 or R ₅ independently replace each of these compounds bifunctional linking group. When R 1 is replaced, it is preferably -CH ₂ -O-CH ₂ - or -CH ₂ -S-CH ₂ C (O) -O- (CH ₂ ) ₂ -OC (O) CH ₂ -S-CH ₂ -.

The invention also relates to compounds of formula I wherein R 1 is - (CH ₂ ) _n - (X) - (O, ⁿ is an integer from 1 to 4, preferably 1, X is oxy, thio or -N (R _a ) preferably thio, Y represents a 2 to 15-membered branched or unbranched carbon chain, optionally containing 1, 2 or 3 non-peroxide oxy, thio or -N (R _a ) - , chain is substituted on carbon with 1, 2 or 3 substituents selected from oxo, carboxy, mercapto, -N (R _b) (R _c) C 3 -C 6 cycloalkyl, aryl, heteroaryl, a saccharide residue or the amino group of the peptide and is saturated or unsaturated, preferably Y represents a 2 to 6-membered branched or unbranched carbon chain, substituted on the carbon atom by 1, 2 or 3 substituents from the group oxo, heteroaryl, amino acid or peptide residue, R 2 represents hydrogen or C 1 -C 6 alkyl, R ₃ represents a hydrogen atom, C 1 -C 6 alkyl, C 1 -C 6 alkoxy, C 1 -C C6alkylthioskupinu, aryl, heteroaryl, aryloxy or heteroaryloxy; R ₄ is hydrogen or C1 -C6 alkyl and R ₅ is hydroxy, C 1 -C 6 alkoxy or C 1 -C C6alkanoyloxyskupinu or consisting of R ₄ and R ₅ together ethylenedioxy, _R6 is hydrogen, carboxy, C 1 -C 6 alkanoyl, C 1 -C 6 alkoxycarbonyl, halogenC 1 -C 6 alkyl, -C (= O) NR _f R _g , a saccharide, amino acid or peptide residue or C 1 -C 6 alkyl optionally substituted with 1 or 2 substituents from hydroxy, C 1 -C 6 alkyl; C 6 alkoxy, C 1 -C 6 alkanoyloxy, carboxy, amino acid, peptide or carbohydrate residue or -C (= O) NR _f R _g , R _a represents a hydrogen atom, C 1 -C 6 alkyl, C 1 -C 6 alkanoyl, phenyl or benzyl and R _b , R _c , R f and R _g independently represent a hydrogen atom of a C 1 -C 6 alkyl, C 1 -C 6 alkanoyl, phenyl or benzyl or form R _b and R _c or R _f and R _g together with the nitrogen atom to which they are attached a pyrrolidine, piperidine or morpholine ring, aryl, heteroaryl, aryloxy or heteroaryloxy of Y or R ₃ is optionally substituted with 1, 2 or 3 substituents from C 1 -C 6 alkyl, C 1 -C 6 alkoxy, C 1 -C 6 alkanoyl, C 1 -C 6 alkanoyloxy, C 1 -C 6 alkoxycarbonyl, hydroxyC 1 -C 6 alkyl, halogenC 1 -C 6 alkyl -C6alkyl, hydroxy, halogen, carboxy, mercapto, nitro or φ «♦♦ · · · · ·φ ·φφφφφφφφφφφφ

-N (Rh) (R j) wherein R _h and R j independently represent hydrogen, C 1 -C 6 alkyl, C 1 -C 6 alkanoyl, phenyl or benzyl or form together with the nitrogen atom to which they are attached a pyrrolidine, piperidine or morpholine ring, as well as pharmaceutically acceptable salts thereof.

The invention also relates to compounds of formula Ii

where

R2 is C1-C6alkyl,

R ₃ is hydrogen, C 1 -C 6 alkyl, C 1 -C 6 alkoxy, C 1 -C C6alkylthioskupinu, aryl, heteroaryl, aryloxy or heteroaryloxy,

R 4 represents a hydrogen atom or C 1 -C 6 alkyl;

R ₅ is hydroxy, C 1 -C 6 alkoxy or C 1 -C 6 alkanoyloxy or R ₄ and R ₅ together form ethylenedioxy,

R ₆ is hydrogen, carboxy, C 1 -C 6 alkanoyl, C 1 -C 6 alkoxycarbonyl, haloC 1 -C 6 alkyl, -C (= O) NR _f R _g , a carbohydrate, amino acid or peptide residue or C 1 -C 6 alkyl optionally substituted with 1 or 2 hydroxy substituents, C1-C6alkoxy, C1-C6alkanoyloxy, carboxy, amino acid, peptide or carbohydrate residue, or -C (= O) NR _f R _g ,

R ₇ is carboxy, C 1 -C 6 alkanoyl, C 1 -C 6 alkoxycarbonyl, haloC 1 -C 6 alkyl, -C (= O) NR _d R _e , a carbohydrate, amino acid or peptide residue, or C 1 -C 6 alkyl, optionally substituted with 1 or 2 substituents from hydroxy, C 1 -C6alkoxy, C1-C6alkanoyloxy, carboxy, amino acid, peptide or carbohydrate, or -C (= O) NR _d R _e ,

Rd. R e, R f and R g independently represent hydrogen, C 1 -C 6 alkyl, C 1 -C 6 alkanoyl, phenyl or benzyl or form R _d and R _e or R f and R _g together with the nitrogen atom to which the pyrrolidine, piperidine or morpholine ring is attached; any aryl, heteroaryl, aryloxy or heteroaryloxy group R ₃ is optionally substituted with 1, 2 or 3 substituents from C 1 -C 6 alkyl, C 1 -C 6 alkoxy, C 1 -C 6 alkanoyl, C 1 -C 6 alkanoyloxy, C 1 -C 6 alkoxycarbonyl, hydroxyC 1 -C 6 alkyl, halogenC 1 hydroxy, halogen, carboxy, mercapto, nitro or -N (Rh) (Rj), wherein Rh and Rj independently represent hydrogen, C1-C6alkyl, C1-C6alkanoyl, phenyl or benzyl or form together with the nitrogen atom to which they are bonded pyrrolidine, piperidine or morpholine ring, as well as pharmaceutically acceptable salts thereof.

The compounds of the present invention are antineoplastic agents, i.e., agents capable of inducing the inhibition of tumor cell growth in vitro or in vivo in mammals, for example humans or domestic animals, and are particularly effective against solid tumors and tumors resistant to a wide variety of active agents. Thus, they inhibit growth of the cells.

The compounds of the invention can be targeted to a particular tumor by combining them with a reagent capable of binding to the tumor-associated antigen. This antigen may be located directly in the tumor or in the area of tumor cells. Suitable reagents in this case are those in this case. polyclonal and monoclonal antibodies. The compound and reagent complex or conjugate of these components may further comprise a linker, for example of the above type, for coupling the active agent to the reagent. Accordingly, the present invention also provides a compound comprising a compound of formula (I) or (II) and a reagent such as a polyclonal or monoclonal antibody capable of binding to an antigen associated with tumor tissue.

Pharmaceutical compositions, eg, single dose dosage forms of one or more compounds of the invention, in combination with a pharmaceutically acceptable diluent or carrier, are also part of the invention.

The present invention also provides compounds of formulas I or II, dimers and conjugates thereof, and a reagent capable of binding to a tumor-associated antigen, or a salt thereof, for use in therapy, preferably for the treatment of cancer, for example solid tumors and the use of said substances for the manufacture of pharmaceutical compositions for said purpose.

The present invention also provides a process for the preparation of said compounds and novel intermediates useful in the preparation of said compounds. Certain compounds of the invention may be used to produce other compounds of the invention.

The invention will now be described with reference to the accompanying drawings.

BRIEF DESCRIPTION OF THE DRAWINGS

Figure 1 shows representative compounds 1 to 9 of the invention and the corresponding intermediates 10 and 11.

9

9

9

9

99 • 9 ··

9 9 · · ··

9 9 9

99

Figure 2 shows representative compounds 13 to 16 of the invention.

Figure 3 shows representative compounds 17 and 18 according to the invention.

In the following description, a halogen atom means a fluorine, chlorine, bromine or iodine atom. Alkyl, alkoxy, alkenyl and the like may have a straight or branched chain, but in the case of individual residues such as propyl, the name includes only a straight-chain residue, a branched-chain isomer such as isopropyl is always specifically mentioned. Aryl is a phenyl or carbon ring bicyclic radical condensed in the ortho position and containing from 9 to 10 ring atoms, at least one of which is aromatic. Heteroaryl includes residues joined by a carbon atom of a monocyclic aromatic ring having 5 or 6 ring atoms, the residue containing carbon atoms and 1 to 4 non-peroxide oxygen, sulfur or N (R _y ) groups where R _y is absent or hydrogen or oxygen , C 1 -C 4 alkyl, phenyl or benzyl, may also be ortho-fused bicyclic, heterocyclic, C 8 -C 10 heterocyclic moieties and especially derived from the benzene derivative or derivative formed by the condensation of a propylene, trimethylene or tetramethylene divalent group with this ring .

The term "inhibition" refers to a decrease in the rate of growth of tumor cells and / or a decrease in tumor weight. Inhibition also includes complete tumor regression. That is, the compounds of the invention may be cytostatic or cytotoxic to tumor cells.

0 · 00

000 ·· ··

0 0

0 0 · 0 0

The compounds of the invention can be administered to mammals suffering from tumors against which the compounds of the invention are effective. The compounds are effective in vivo as well as in vitro on human tumor cell lines. They are particularly suitable for the treatment of solid tumors for which insufficient chemotherapeutic agents are currently available. Thus, the compounds of the invention are effective, e.g., against epidermoid and myeloid tumors in acute (AML) or chronic (CML) form. These tumors also include tumors other than small cell tumors, liver tumors, tumors in the cervical region, kidneys, adrenal, stomach, esophagus, oral cavity, mucosal and skin tumors, as well as lung, ovarian, mammary, colon and melanoma tumors. including amelanotic subtypes. In addition, the compounds may also be used for cervical, bladder, pancreatic, lymphoma, Hodgkin's disease, prostate, sarcoma and testicular tumors, as well as for tumors of the central nervous system such as brain tumors, neuroblastomas and hematopoietic stem cell tumors such as B cell leukemia / lymphomas, myelomas, T cell leukemia / lymphomas, small cell leukemia / lymphomas, monocytic leukemia, myelomonocytic leukemia, and Hairy cell leukemia. These leukemias / lymphomas may be acute (ALL) or chronic (CLL).

The term carbohydrate refers to monosaccharides, disaccharides, trisaccharides and polysaccharides. Thus, the term includes glucose, sucrose, fructose and ribose as well as deoxy sugars such as deoxyribose and the like. Carbohydrate derivatives are readily prepared by the method described in International Patent Applications WO 96/34005 and WO 97/03995. The saccharide can be readily attached to the remainder of the compound of formula (I) or (II) by an ether bond.

Amino acid means natural amino acid residues such as Aia, Arg, Asn, Asp, Cys, Glu, Gln, φ • φ φ φ φ φ φ φ φ φ φ φ φ φ · · · · · φ φ • φ • φ φφφ φφφ φ

φ φ

φ

Gly, His, Hyl, Hyp, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, and Val in the D- or L-form as well as non-natural amino acid residues such as phosphoserine, phosphothreonine, phosphotyrosine, hydroxyproline, gamma-carboxyglutamate, hippuric acid, octahydroindole-2-carboxylic acid, statin, 1,2,3,4-tetraisoquinoline-3-carboxylic acid, penicillamine, ornithine, citrulline, alpha-methylalanine, p-benzoylphenylalanine , phenylglycine, propargylglycine, sarcosine and tert-butylglycine. The term also encompasses both natural and non-natural amino acids having a conventional protecting group such as an acetyl or benzyloxycarbonyl group on an amino group or having a protecting group on a carboxyl group such as a C 1 -C 6 alkyl group or a phenyl or benzyl ester group. Other suitable amino and carboxyl protecting groups are well known in the art, and are disclosed, for example, in TW Greene, Protecting Groups In Organic Synthesis, Wiley: New York 1981. The amino acid may be bound to the remainder of a compound of Formula I by its carboxyl, amino group. or by any other means, for example via a cysteine sulfur atom.

By peptide is meant a sequence of 2 to 25 amino acid residues as defined above, or any peptide residue. The sequence used may be linear or cyclic. The cyclic peptide can be prepared by forming disulfide bridges between two cysteine residues in the sequence. The peptide moiety may be attached to the remainder of the molecule of formula (I) by carboxyl, amino or any other suitable means, for example via a cysteine sulfur atom. Preferably, the peptide comprises 3 to 25, and in particular 5 to 21 amino acid residues. Peptide derivatives may be prepared as described in US 4612302, US 4853371 and US 4684620 or as described in the following examples. Specifically reported peptide sequences are shown with p

0

0 «··

0

0

0

000 000 · ♦ «

0 «« · · 0 · · ·

0 0 ·

0 0 · ·· ·· amino terminal on the left side and carboxyl group on the right side.

It has been shown that some peptides can specifically bind to specific tumor-associated antigens in a manner similar to the binding of antibodies to these antigens, as described, for example, in Arap et al., Science, 1998, 279, 5349, 377-380 . In this way, it is preferable to contact a peptide with said ability, i.e., the ability to bind to specific receptors on tumor cells towards tumor tissue. Accordingly, a preferred embodiment of the invention is a compound of formula I or II containing the peptide residue, capable of specifically binding to a tumor-associated antigen.

Preferred peptides are, for example, -Cys-Asp-Cys-Arg-Gly-Asp-Cys-Phe-Cys (SEQ ID NO: 1), -Cys-Asp-Gly-Arg-Cys (SEQ ID NO: 2), and - Cys-Asp-Gly-Cys-Lys-Asn-Phe-Phe-Trp-Lys-Thr-Phe-Thr-Ser-Cys (SEQ ID NO: 3).

It will be appreciated that compounds of the invention that contain a center of chirality may exist and may be isolated in optically active and racemic forms. Some of these may be polymorphic. It is to be understood that the invention encompasses any racemic, optically active, polymorphic or stereoisomeric forms of the compounds of the invention or mixtures thereof with the desirable properties described. It is known to prepare optically active forms, for example, by resolution of the racemic form by recrystallization, synthesis from optically active starting materials, chiral synthesis or chromatographic separation using a chiral stationary phase. Next, it will be described how the antitumor activity of these compounds can be determined using Test A or Test B described below, or other known tests. In preferred compounds of Formula I, the absolute stereochemistry at the carbon atom bearing R 4 and R 5 is (R).

Flf fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl Fl.

The specific values given below for individual residues or substituents are illustrative only. Thus, they do not preclude the use of other radicals as defined above for individual radicals or substituents.

C1-C4alkyl refers to methyl, ethyl, propyl, isopropyl, butyl, isobutyl or sec-butyl, C1-C6alkyl refers to methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, pentyl, 3- pentyl, or hexyl, C 1 -C 8 alkyl may be methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, pentyl, 3-pentyl, hexyl, heptyl or octyl, C 3 -C 6 cycloalkyl may be cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl C1-C4alkoxy may be methoxy, ethoxy, propoxy, isopropoxy, butoxy, isobutoxy or sec.butoxy, C1-C6alkoxy may be methoxy, ethoxy, propoxy, isoxy, isopropoxy, isopropoxy, isopropoxy, isopropoxy, isopropoxy,

3-pentoxy or hexyloxy, C2-C6alkenyl means vinyl, allyl, 1-propenyl, 2-propenyl, 1-butenyl, 2-butenyl, 3-butenyl, 2-methyl-2-propenyl, 1-pentenyl, 2-pentenyl, 3-pentenyl, 4-pentenyl, 3-methyl-2-butenyl, 3-methyl-3-butenyl, 1-hexenyl, 2-hexenyl, 3-hexenyl,

4-hexenyl, 5-hexenyl or 4-methyl-3-pentenyl, C 1 -C 6 alkanoyl may be acetyl, propanoyl or butanoyl, the halo C 1 -C 6 alkyl may be iodomethyl, bromomethyl, chloromethyl, fluoromethyl, trichloromethyl, trifluoromethyl, 2-chloroethyl, 2 -fluoroethyl, 2,2,2-trifluoroethyl or pentafluoroethyl, the hydroxyC 1 -C 6 alkyl may be hydroxymethyl,

1-hydroxyethyl, 2-hydroxyethyl, 1-hydroxypropyl, 2-hydroxypropyl,

3-hydroxypropyl, 1-hydroxybutyl, 4-hydroxybutyl, 1-hydroxypentyl,

5-hydroxypentyl, 1-hydroxyhexyl or 6-hydroxyhexyl,

C 1 -C 6 alkoxycarbonyl may be methoxycarbonyl, ethoxycarbonyl, propoxycarbonyl, isopropoxycarbonyl, butoxycarbonyl,

Pentoxycarbonyl or hexyloxycarbonyl, C 1 -C 6 alkylthio may be methylthio, ethylthio, propylthio, isopropylthio, butylthio, isobutylthio, pentylthio or hexylthio, acetyloxy, , butanyoloxy-, isobutanoyloxy-, pentanoyloxy-, or hexanoyloxy, the aryl may be phenyl, indenyl or naphthyl and the heteroaryl may be furyl, imidazolyl, triazolyl, triazinyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, pyrazolyl, pyrrolyl, pyrolyl, pyrolyl, pyrolyl, pyrolyl, pyrolyl, pyrolyl, pyrolyl or an N-oxide, thienyl, pyrimidinyl or N-oxide, indolyl, isoquinolyl or N-oxide or quinolyl or N-oxide thereof.

A specific value for R 1 is hydroxy, halogen, carboxy, aryl, heteroaryl and a saccharide, amino acid or peptide residue.

Another specific value for phy is - (CH ₂ ) _n - (X) - (Y), wherein n = 0 to 4, X is oxy, thio, -N (R _a ) - or is absent and Y is the remainder of the peptide or C 1 -C 6 alkyl, substituted with a peptide residue.

Another specific value for R 1 is a hydrogen atom, or C 1 -C 6 alkyl, optionally substituted with 1 or 2 substituents selected from hydroxy, halogen, methoxy or ethoxy.

Another specific value for R 1 is - (CH ₂ ) _n - (X) - (Y) wherein X is oxy, thio, or -N (R _a ) - and Y is CH ₂ OC (O) (C 1 -C 4 alkyl) ), CH ₂ C (O) - (CH ₂ ) ₂ -OC (O) CH ₂ SH, (CH ₂ ) ₂ -O- (CH ₂ ) ₂ halide, (C 1 -C 4 alkyl) -O- (C 1 -C 4 alkyl) CH ₂ CO ₂ (C 1 -C 4 alkyl), CH ₂ CO ₂ H, arylC 1 -C 5 alkyl, a saccharide or amino acid residue or C 1 -C 8 alkyl, optionally substituted with 1 or 2 hydroxy or halogen atoms, wherein any aryl

9

8 4

4,444 4

4

4 4 4 4

98 884 «88 44

8 9 4 8 9

8 4 4 4

4 9 4 8 8

448 84 48 or a heteroaryl group of Y is optionally substituted with 1 or 2 substituents selected from hydroxy, halogen, C 1 -C 4 alkyl or C 1 -C 4 alkoxy.

Another specific value for R 1 is -CH 2 - (X) - (Y) wherein X is oxy, thio or -N (R _a ) - and Y is C 1 -C 8 alkyl, optionally substituted with 1 or 2 substituents from hydroxy, atom halogen, carboxy, oxo, mercapto, -N (R b) (R _c ), C 3 -C 6 cycloalkyl, aryl, heteroaryl, saccharide, amino acid or peptide moiety, wherein any aryl or heteroaryl group of Y is optionally substituted, 2 or 3 C1-C6 alkyl, C1-C6 alkoxy, C1-C6alkanoyl, C1-C6alkanoyloxy, C1-C6alkoxycarbonyl, hydroxyC1-C6alkyl, haloC1-C6alkyl, hydroxy, halogen, mercapto, nitro or -N (Rh) (Rj) substituents wherein R h and R j independently represent hydrogen, C 1 -C 6 alkyl, C 1 -C 6 alkanoyl, phenyl or benzyl or form R h and R j together with the nitrogen atom to which they are attached a pyrrolidine, piperidine or morpholine ring.

Another specific value for R 1 is - (CH ₂ ) _n - (X) - (Y) wherein n = 1 or 2, X is oxy, thio or -N (R _a ) - and Y is C 1 -C 6 alkyl or C 2 -C 6 alkenyl, optionally substituted with 1 or 2 substituents from oxo, hydroxy, carboxy, halogen, mercapto, nitro, -N (R _b ) (R c), C 3 -C 6 cycloalkyl, aryl, heteroaryl, saccharide, amino acid or peptide radical, wherein any aryl or heteroaryl group of Y is optionally substituted with 1, 2 or 3 substituents from C 1 -C 6 alkyl, C 1 -C 6 alkoxy, C 1 -C 6 alkanoyl, C 1 -C 6 alkanoyloxy, C 1 -C 6 alkoxycarbonyl, hydroxyC 1 -C 6 alkyl, halogenC 1 -C 6 alkyl, hydroxy, halogen, mercapto, nitro or -N (Rh) (Rj), wherein Rh and Rj are independently hydrogen, C1-C6alkyl, C1-C6alkanoyl, phenyl;

99 «

9

9 9 • 9

99 9 9 9

9 9 ·

9 9 9 ·

Or benzyl or forms R _h and R j together with the nitrogen atom to which they are attached a pyrrolidine, piperidine or morpholine ring.

Another specific value for R 1 is -CH ₂ - (a sulfur-bound cysteine residue) -R _x , wherein R _x is an amino acid or peptide residue of 2 to 24 amino acid residues.

Another particular value for R is -CH ₂ - (N-acylcysteines radical bonded via a sulfur atom) -R _x wherein R _x is an amino acid residue or a peptide containing 2-24 amino acid residues.

Another specific value for Rt is -CH ₂ - (glutathione, linked via a sulfur atom).

Another specific value for Rt is 3 - [(R) -alpha-methylbenzylaminocarbonyl] -2- (acylamino) ethylthiomethyl. The ethyl group at the 2-position preferably has the (S) -configuration of cysteine.

A preferred value for Rt is hydrogen, methyl, hydroxymethyl, methoxymethyl or acetoxymethyl.

A particular value for _R2 is hydroxymethyl, methoxymethyl, or acetoxymethyl.

Another specific value for R ₂ is carboxy, C 1 -C 6 alkanoyl, C 1 -C 6 alkoxycarbonyl or -C (= O) NR d R _e .

Another specific value for R ₂ is C 1 -C 6 alkyl, substituted with 1 or 2 substituents selected from hydroxy, C 1 -C 6 alkoxy, C 1 -C 6 alkanoyloxy, carboxy, amino acid residue, peptide or carbohydrate, or -C (= O) NR d R _e . · · · · · · · · · · · · · · · · · · · · · ·

Another specific value for R ₂ is -CH ₂ - (a sulfur-bound cysteine residue) -R _x , wherein R _x is an amino acid or peptide residue of 2 to 24 amino acid residues.

Another specific value for R ₂ is -CH _{2 -} (a sulfur-bound N-acylcysteine residue) -R _x , wherein R _x is an amino acid residue or a peptide containing 2 to 24 amino acid residues.

Another specific value for R ₂ is -CH ₂ - (glutathione, bonded through a sulfur atom).

A particular value for R ₃ is hydrogen.

A specific value for R4 is methyl.

A specific value for R5 is hydroxy.

A specific value for R ₆ is hydrogen.

Another specific value for R 6 is carboxy, C 1 -C 6 alkanoyl, C 1 -C 6 alkoxycarbonyl or -C (= O) NR _f R _g .

Another specific value for R 6 is C 1 -C 6 alkyl, optionally substituted with 1 or 2 substituents from the group C 1 -C 6 alkoxy, C 1 -C 6 alkanoyloxy, carboxy, amino acid residue, peptides or carbohydrate, or -C (= O) NR _f R _g .

Another specific value for R ₆ is -CH ₂ - (a sulfur-bound cysteine residue) -R _x , wherein R _x is an amino acid or peptide residue of 2 to 24 amino acid residues.

Another specific value for R ₆ is -CH ₂ - (residue

N-acylcysteine, linked via a sulfur atom) -R _x , wherein R _x is an amino acid residue or a peptide of 2 to 24 amino acid residues.

A preferred value for R 6 is methyl or hydroxymethyl.

Of particular interest for Ry is C 1 -C 6 alkyl, substituted with 1 or 2 substituents selected from hydroxy, C 1 -C 6 alkoxy, C 1 -C 6 alkanoyloxy, carboxy, amino acid, peptide or carbohydrate, or -C (= O) NR _d R _e .

A specific value for R ₇ is also hydroxymethyl.

A specific value for R _x is a peptide of 4 to 20 amino acid residues.

Another specific value for R _x is -Leu-Gly-Phe, -Phe-Leu-Gly, -Leu-Leu-Phe, -Gly-Phe, or -Leu.

Another specific value for R _x is -Asp-Cys-Arg-Gly-Asp-Cys-Phe-Cys (SEQ ID NO: 4), -Asp-Gly-Arg-Cys (SEQ ID NO: 5), or -Asp- Gly-Cys-Lys-Asn-Phe-Phe-Trp-Lys-Thr-Phe-Thr-Ser-Cys (SEQ ID NO: 6).

A specific compound of the invention is, for example, a compound of formula I wherein R ₂ is C 1 -C 6 alkyl substituted by hydroxy, C 1 -C 6 alkoxy, C 1 -C 6 alkanoyloxy or carboxy, R ₃ is hydrogen,

C 1 -C 6 alkoxy, C 1 -C C6alkylthioskupinu, aryl, heteroaryl, aryloxy or heteroaryloxy; R ₄ is hydrogen or C1-C6 alkyl, R is hydroxy and R6 is C1 -C6 alkyl optionally substituted with hydroxy, • ···· oo · · · · · <

C1-C6alkoxy, C1-C6alkanoyloxy or carboxy and pharmaceutically acceptable salts thereof.

Another specific compound of the invention may be a compound of formula I wherein Rf is hydrogen or C1-C6 alkyl optionally substituted by hydroxy, halogen, methoxy, ethoxy or acetoxy,

R ₂ is hydroxymethyl, methoxymethyl, or acetoxymethyl; R ₃ is hydrogen, _R4 is methyl, _R5 is hydroxy and R6 is methyl or hydroxymethyl, and pharmaceutically acceptable salts thereof.

Another specific compound of the invention is a compound of formula I wherein R 1 is - (CH ₂ ) _n - (X) - (Y), n = 0 to 4, X is oxy, thio, -N (R _a ) -, or is absent, Y represents an amino acid residue with 1 or 2 protecting groups and R ₂ represents a hydrogen atom or C 1 -C 6 alkyl, as well as pharmaceutically acceptable salts thereof.

Another specific compound of the invention is a compound of formula I wherein R 1 is -CH ₂ - (a N-acylcysteine moiety bound through a sulfur atom), (S) -2 - [(R) -alpha-methylbenzylaminocarbonyl] -2- (acylamino) ) ethylthiomethyl or (R) -2 - [(R) -alpha-methylbenzylaminocarbonyl] -2- (acylamino) ethylthiomethyl and R ₂ represents a hydrogen atom or C 1 -C 6 alkyl, as well as pharmaceutically acceptable salts thereof.

A specific compound of formula I is a compound of formula

O

9

9 9 9 9

Or a pharmaceutically acceptable salt thereof.

A specific compound of formula II is a compound of formula

or a pharmaceutically acceptable salt thereof.

A preferred compound of the invention is a compound of formula I wherein R ₄ is hydrogen or C 1 -C 6 alkyl and R 5 is hydroxy or acetoxy, wherein the absolute stereochemistry at the carbon atom bearing R ₄ and R ₅ is (R) as well as pharmaceutically acceptable salts thereof.

The compounds of the present invention can be prepared by the methods described hereinafter, with the substituents as defined above, unless otherwise specifically indicated for some substituents.

Compounds of the invention, i.e. compounds of formula I and II and their salts may be derived from illudin S, hydroxymethylacylfulvene HMAF (i.e. compound of formula I in which R is CH ₂ OH, R ₂ is methyl, R ₃ is hydrogen R ₄ is methyl, R 5 is OH and R 5 is methyl) and fulvene (i.e., compounds of formula I wherein R 1 is hydrogen, R ₂ is methyl, R ₃ is hydrogen, R ₄ is methyl, R ₅ is OH and R6 are

• 4 4 4

4 4 4

4 4 4

4 4 4

In the preparation of the compounds according to the invention, known processes are described, for example, in WO 91/04754 and WO 94/18151.

A compound of formula I wherein R ₂ is hydroxymethyl may be prepared by oxidising a corresponding compound of formula I wherein R ₂ is methyl. Oxidation is readily accomplished by treatment with selenium dioxide and tert-butyl hydroperoxide under conditions similar to those described in Example 1.

The compound of formula I wherein R 1 is hydroxymethyl can be prepared from the corresponding compound of formula I wherein R 1 is hydrogen by treatment with paraformaldehyde and sulfuric acid. The reaction proceeds well under the conditions described in Example 2.

A compound of formula I wherein _R2 is acetoxymethyl can be prepared by acylating the corresponding compound of formula I wherein R ₂ is hydroxymethyl. Acylation can be accomplished with acetic anhydride under the conditions described in Example 3.

A compound of formula I wherein _R2 is methoxymethyl can be prepared by reacting the corresponding compound of formula I wherein _R2 is hydroxymethyl with methyl iodide and silver oxide. The reaction may be carried out under the conditions of Example 7.

A compound of formula I wherein Rf is methoxymethyl may be prepared by reacting a corresponding compound of formula I wherein R1 is hydroxymethyl with methanol and sulfuric acid. The reaction proceeds well under the conditions described in Example 8, part (a).

A compound of formula I wherein R is -CH ₂ - (a cysteine residue, bonded via a sulfur atom) is obtained by reacting a corresponding compound • • ·· · · · · · • «· · ·

Formula I wherein R 1 is hydroxymethyl, with cysteine. The reaction may be carried out under the conditions of Example 10.

A compound of formula I wherein R 1 is -CH ₂ - (a sulfur-bound N-acylcysteine residue) can be prepared by reacting the corresponding compound of formula I wherein R 1 is hydroxymethyl with N-acylcysteine. The reaction can be easily carried out under the conditions of Example 11.

A compound of formula I wherein R 1 is 2 - [(R) -alpha-methylbenzylaminocarbonyl] -2- (acylamino) ethylthiomethyl can be prepared by reaction of a corresponding compound of formula I wherein R 1 is -CH ₂ - (N-acylcysteine residue) bonded via a sulfur atom) with alpha-methylbenzylamine. The reaction proceeds well under the conditions of Example 12.

A compound of formula I wherein R 1 is -CH ₂ - (a sulfur bonded glutathione moiety) can be prepared by reacting the corresponding compound of formula I wherein R 6 is hydroxymethyl with glutathione. The reaction proceeds readily under the conditions of Example 14.

A compound of formula I wherein R 1 is -CH ₂ - (sulfur-bonded cysteine) -R _x or CH ₂ - (sulfur-bonded N -acylcysteine) -R _x may be prepared by preparing: reacting the corresponding compound of formula I wherein R 1 is -CH ₂ - (a sulfur-linked cysteine residue) or CH ₂ - (a sulfur-bound N-acylcysteine residue) with an appropriate amino acid or peptide corresponding to R _x . The reaction proceeds readily under the conditions of Example 15.

• 9 • 9 9 · 9 9 · ··· ·

9 9 9 9 9 • 999 8

99 999

The compound of formula (II) can be prepared from illudine S by the method of Example 17.

Compounds of the invention may also be prepared using the procedures and intermediates disclosed in T. McMorris et al., Tetrahedron, 1997, 53, 44, 14579-14590, T. McMorris et al., J. Org. Chem. 1997, 62, 3015-3018, T. McMorris et al., Chem. Comm. 1997, 315-316, T. McMorris et al., Experientia, 1996, 52, 75-80 and also in US 5439942, US 5439936, US 5523490, US 5536176, WO 91/04754, WO 94/18151 and WO 98 / 03458. Certain compounds of formulas I and II may be used to prepare other compounds of formulas I or II.

If the compounds obtained are sufficiently basic or acidic to form stable, non-toxic salts with acids or bases, it may be appropriate to administer the compounds in the form of salts. Examples of pharmaceutically acceptable salts include organic acid addition salts forming a physiologically acceptable anion, such as tosylate, methanesulfonate, acetate, citrate, malonate, tartrate, succinate, benzoate, alpha-ketoglutarate and alpha-glycerophosphate. Inorganic salts include hydrochlorides, sulfates, nitrates, hydrogen carbonates and carbonates.

Pharmaceutically acceptable salts may be prepared using conventional methods known in the art, for example by reacting a sufficiently basic compound, for example an amine, with a suitable acid to form a physiologically acceptable anion. In the case of carboxylic acids, alkali metal salts such as sodium, potassium or lithium salts or alkaline earth metal salts such as calcium salts may be formed.

The compounds of the present invention may be formulated into pharmaceutical compositions and administered to mammals in such form, for example, orally or parenterally, to a human in a dosage form adapted to the route of administration selected. , intravenously, intramuscularly, topically or subcutaneously.

The compounds of the invention may be administered systemically, for example orally, in combination with a pharmaceutically acceptable carrier, for example, an inert diluent or an absorbable edible carrier. These materials may be administered in hard or soft gelatin capsules, compressed into tablets, or incorporated into a patient's diet. For oral administration, the active ingredient is usually mixed with one or more excipients and then taken in the form of tablets, oral mucosa, capsules, elixirs, suspensions, syrups, wafers and the like. Said compositions should contain at least 0.1% of active ingredient. However, the active ingredient content of these compositions can be varied and is usually in the range of 2 to 60% by weight of the individual dosage forms. The amount of active ingredient should always be such that an effective concentration in the bloodstream is achieved.

Tablets, pills, capsules, and the like may contain various excipients such as binding agents such as gum tragacanth, acacia, corn starch or gelatin, bulking agents such as calcium hydrogensophate, a disintegrating agent such as corn starch, potato starch, alginic acid and the like. likewise, glidants such as magnesium stearate and sweeteners such as sucrose, fructose, lactose or aspartame or else flavorings such as mint oil or cherry flavor. When the dosage form used is a capsule, the capsule may contain, in addition to materials of the above type, a liquid carrier such as a vegetable oil or polyethylene glycol. Various other materials may be used as coatings or to otherwise modify the physical properties of the solid dosage form. For example, tablets, pills, or capsules may be provided with 9

9 9 ·· • · ·

9 9 9 9 9 9 9 9 9 9 9 9 9 9 9 9 9 9 9 9 9 9 9 9 9 9 9 9 9 9 9 9 9 9 , sugar, and the like. Syrups or elixirs may contain the active ingredient, sucrose or fructose as a sweetening agent, methyl and propylparabens as a preservative, and in addition a coloring and flavoring agent, for example cherry orange flavor. Of course, any material used to make the dosage form should be pharmaceutically acceptable and substantially non-toxic in the amount used. In addition, the active ingredient may be formulated to be delayed or delayed release.

The active compounds may also be administered intravenously or intraperitoneally by infusion or injection. Solutions of the active ingredient or a salt thereof may be prepared in water, optionally mixed with a nontoxic surfactant. Dispersions can also be prepared in glycerol, liquid polyethylene glycols, triacetin and mixtures thereof, as well as in oils. Under normal conditions of storage and use, these compositions will contain preservatives to prevent the growth of microorganisms.

Pharmaceutical dosage forms for administration by injection or infusion include sterile aqueous solutions or dispersions or sterile powders containing the active ingredient intended for the preparation of sterile solutions or dispersions just prior to use, the active ingredient optionally being encapsulated in liposomes. In all cases, the resulting dosage form must be sterile, fluid and stable under the conditions of manufacture and storage. The liquid carrier may be a solvent or a liquid dispersion medium such as water, ethanol or polyol, e.g., glycerol, propylene glycol, liquid polyethylene glycols, and the like. Vegetable oils, non-toxic glyceryl esters and mixtures thereof may also be used. The fluidity can be adjusted, for example, by forming liposomes and maintaining the desired particle size in the case of dispersion, or by using a wetting agent. Preservatives may be used

09 · · 90 ·· · 9 · 9 · 8 8 8 8 9 9

9 9 98 9 9 9 9 9

9 9 8 8 9 9 9 9 8 8 9

9 9 9 * 9 9 0 0 9 • 9 99 999 099 Μ 09 various antibacterial and antifungal agents such as parabens, chlorobutanol, phenol, sorbic acid, thimerosal and the like. In many cases, it will be advantageous to add substances to provide the desired osmotic pressure, such as sugars, buffers, or sodium chloride. Prolonged absorption of injectables can be ensured by the inclusion of agents that delay absorption such as monostearate and gelatin.

Sterile injectable solutions are prepared by mixing the active ingredient with the required amount of the appropriate solvent and optionally the above excipients as desired, then sterilizing the mixture by filtration. In the case of sterile powders, the sterile injectable solutions are dried under vacuum and lyophilized to give the active ingredient in the form of a powder, containing the other ingredients present in the dried solution.

For topical administration, the compounds of the invention may be administered in pure form when they are liquids. However, they are usually applied in the form of compositions comprising a dermatologically acceptable carrier, solid or liquid.

Solid carriers include finely powdered solids such as talc, clays, microcrystalline cellulose, silica, alumina and the like. The liquid carrier usable may be water, alcohols or glycols or mixtures of water, alcohol or glycol, in which the active ingredients may be dissolved or dispersed in effective concentrations, optionally in admixture with nontoxic surfactants. For example, fragrances and antimicrobials may be used as adjuvants. The resulting liquid compositions can be applied by cotton wool, used to impregnate dressings, or sprayed, for example, as an aerosol.

99 • 9

99 »· 9 9 9 9 9 9 9

99 • 9 • 9 • 9

9

999 999

9 ·

9 9

9 9 • 9 9

Thickening agents such as synthetic polymers, fatty acids and their salts and esters, fatty alcohols, modified celluloses or modified inorganic materials can also be used in the liquid carrier to give spreadable pastes, gels, lubricants, soaps and the like for direct application to the skin. user. Examples of suitable dermatological compositions by which a compound of the invention can be applied to the skin are generally known and have been described, for example, in US 4608392 (Jacquet et al.), US 4992478 (Geria), US 4559157 (Smith et al.) And US 4820508 ( Wortzman).

Appropriate dosages of the compounds of the invention can be determined by correlating between their in vitro activity and in vivo activity in an animal model, for example, in mice or dogs, known assays developed for illudine analogs and described in US 5439936 and US 5523409. an assay for efficacy in higher mammals including children and adults in US 4938949 (Borch et al.).

The therapeutically effective amount of the compounds of the invention varies with the subject to be treated and the tumor to be treated. It has been found that relatively high doses of representative compounds of the invention can be administered due to reduced toxicity compared to illudin S and M. For example, the maximum tolerated dose of illudin S is approximately 250 micrograms / kg, while compound 2 can be chronically administered at 25 milligrams / kg without toxicity. A dose of 30 to 112000 micrograms / kg body weight is particularly suitable for intravenous administration, while for intraperitoneal administration, an effective dose of 300 to 112000 micrograms / kg body weight is effective. It will be appreciated that the amount of active ingredient used may vary depending upon the route of administration. The amount of the compound or salt or derivative employed will also vary with the particular compound employed, the route of administration, the nature of the condition being treated and the age and general condition of the patient, and the resulting dose will always be determined by the attending physician.

0 * 00

0 0

0 0 0 0

0 0

0 0 0

00

0 «00 00 0 · 0 * 00

0 0 0 0 0

0 0 0 0 0 0 0 0 0 0 0 0 0

000 000 00 00

The compounds of the invention may be conveniently administered in unit dosage forms, for example 5 to 1000 mg, preferably 10 to 750 mg, and especially 50 to 500 mg.

Ideally, the active agent should be administered to achieve maximum plasma concentrations of the active agent in the range of 0.5 to 75 microM, preferably 1 to 50 microM, and especially 2 to 30 microM. This can be achieved, for example, by intravenous injection of a solution of the active compound at a concentration of 0.05 to 5%, optionally in physiological saline or by oral administration of 1 to 100 mg of the active compound. Desirable circulating concentrations may be maintained by continuous infusion of the active ingredient in an amount of 0.01-5.0 mg / kg / h or by intermittent infusion containing 0.4-15 mg / kg of active ingredient.

The desired dose may be administered at once or divided throughout the day at appropriate intervals, which may be 2, 3, 4, or more sub-doses throughout the day.

The cytotoxic and antitumor properties of the active compounds can be determined using commonly known pharmacological models or using tests A and B below.

Test A: In vitro test

In order to detect cytotoxic effects, various concentrations of compounds of the invention were added to a culture of MV522 (human lung carcinoma cell line), HL60 (myeloid leukemia cell) and 8392 (B cell leukemia / lymphoma cells) cultures for 48 hours and then determined. cell growth and cell viability by trypan blue exclusion It is also possible to apply the cells in liquid culture to 96-well plates and expose them to · · φ φ φ φ φ φ φ φ φ

ΦΦ ΦΦ ·· * · ΦΦΦΦ

Φ Φ ΦΦΦΦ

Φ ΦΦΦΦΦΦ

Φ Φ ΦΦΦΦ

ΦΦ ΦΦ ΦΦ different concentrations of the active substance for 2 hours, then add ³ H-thymidine for 1 to 2 hours and transfer the material to filter paper.

Filter papers are added to vials containing scintillation fluid and the radioactivity is measured using a beta counter.

The results of Test A for representative compounds of the invention are shown in Table 1 below. Values are given as mean ± standard deviation, units are nanomoles / L, NT means no experiment was performed.

Table 1

2 h _IC50 (nm / l) Compound MV522 HL60 8392 1 2640 ± 360 NT 37000 ± 2300 2 11300 ± 1500 NT NT 3 19600 ± 9700 15600 ± 4600 62000 ± 3600 4 20400 ± 6300 > 40000 NT 5 24000 ± 6100 38,400 ± 9,000 NT 6 NT NT NT 7 7700 ± 3500 6000 ± 1200 NT 8 > 80000 > 80000 NT 9 > 50000 49800 ± 20000 NT - 13 10000 ± 1800 NT NT 14 3050 ± 550 NT NT 15 Dec NT NT NT

Test B: In vivo test

Several representative compounds of the invention were used to perform the in vivo assay. The antitumor substance mitomycin C was used as a control. The active substances were administered from day 10 after inoculation daily intraperitoneally for 5 consecutive times.

0 0 0 0 0

0 000 0

0 0 0 0 0

0 0 0 0

00 000 ·

00

0 0 0

0 0 0

0 0 0 • 0 0 0

00 following days. The animals were followed for three weeks from the start of treatment. The MTD was achieved for the control substance mitomycin C, but not for compounds 1 or 2.

4-week-old female BALB / c nu / nu strains weighing 18-22 g were obtained from Simonsen Inc. (Gilroy CA) and maintained in an athymic mouse colony at the University of California, San Diego CA in the absence of pathogenic organisms in HEPA filter cages. Animals were given sterilized food and water ad libitum. The animals were housed 5 in plastic cages ventilated through polyester filters. The animal handling staff used clean sterilized coats, gloves, face masks, headgear and shoes. All experiments were conducted according to the NIH guidelines of the University Institutional Animal Care and Use Committee (Protocol 3-006-2).

The MV522 lung carcinoma cell line was derived by Kelner et al., Anticancer Res., 15: 867-872 and 873-878, 1995 and maintained in an antibiotic-free RPMI 1640 environment (Mediatech, Herndon, VA) supplemented with 10% fetal calf serum. and 2 mM glutamine in a humidified carbon dioxide incubator maintained at 37 ° C.

Mice were randomized into groups of five animals in the initial experiments and groups of 16 to 20 animals to confirm the efficacy of the analog. Each animal was labeled on the ear and monitored individually throughout the experiment. Mice were injected with 10 million MV522 cells per injection in the shoulder region. After ten days, the subcutaneous tumors were approximately 3x3 mm in size and the animals received the active substance. Thereafter, the effect of the active ingredient on prolonging life was monitored.

• · · ·

9 · * 9

9 9 9

9 9 9 • 9 ♦

99

9 9 * 9 9 9

9

9 9 9

99

Although the MV522 cell line induces metastatic death in mice, primary tumor growth from the first day of treatment was also monitored at weekly intervals. Tumor size was measured in two perpendicular directions. Tumor weight was determined by the mass = (width) ² x length / 2 equation. The relative weight of RW was calculated to standardize tumor size variability in the initial treatment groups using the formula RW = Wt / Wi, where Wi is the tumor weight of the animal at the start of treatment and Wt is the tumor weight after some time from the start of treatment. The animals were sacrificed after some time and the organs were monitored for the presence of metastases.

Kaplan and Meier animal survival curves were used for comparison. Conventional ANOVA followed by multiple Tukey-Kramer comparisons according to Kelner et al., Anticancer Res., 15: 867-872 and 873 and 878, 1995 were used to compare the relative tumor weights between groups of animals. Values were statistically significant at (p) <0.05.

Test B data for representative compounds of the invention are shown in Table 2. 8 animals in the control group and 4 animals for each treatment group were used. The values indicated represent the relative weight of the tumor, i.e. Wt / Wi (at day 10 this value was 1.0). The active ingredients were administered intraperitoneally on days 10, 11, 12, 13 and 14 (QD x 5 days).

• 44

4 9 9 9 9 9 4

9 444 9 9 4 4 4 4 • 9 9 4 9 4

9 4 4 4 9 · «« · · 94 94 444 94 49

Table 2

Compound Den 17 Day 24 Den 31 Control (physiological NaCl solution) 4.3 ± 0.7 7.0 ± 1.3 11.8 ± 3.0 1 (15 mg / kg) 1.4 ± 0.7 7.0 ± 1.3 11.8 ± 3.0 2 (35 mg / kg) 2.1 ± 0.2 3.3 ± 0.3 4.0 ± 0.9 13 (32 mg / kg) 2.3 ± 0.5 4.1 ± 0.8 4.7 ± 1.5 14 (28 mg / kg) 3.2 ± 0.6 4.1 ± 0.8 6.0 ± 1.8

The data of Tables 1 and 2 show that representative compounds of the invention are potent cytotoxic and antitumor agents.

The invention will be further illustrated by the following examples, which are not intended to limit the scope of the invention. In the example, melting points are reported without correction, ¹ H and ¹³ C NMR spectra were measured at 300 and 75 MHz, high resolution mass spectra were determined at the University of Minnesota Mass Spectrometry Service Laboratory, Chromatography was performed on Davisil silica gel sized 230-425 mesh (Fisher Scientific) eluting with ethyl acetate / hexanes, TLC analytical thin-layer chromatography was performed on Whatman 4420 222 silica gel plates, and reactions were routinely monitored by TLC. Illudine S, hydroxymethylacylfulven HMAF and fulven were prepared as described above, for example according to international patent applications WO 91/04754 and WO 94/18151.

DETAILED DESCRIPTION OF THE INVENTION

Example 1

Compound 1 (a compound of formula I wherein R 1 is hydrogen, R ₂ is hydroxymethyl, R ₃ is hydrogen, R ₄ is methyl, R 5 is hydroxy and R ₆ is methyl.

Flfl flfl flfl flfl flfl flfl fl flfl fl flfl flfl flfl flfl flfl flfl flfl flfl flfl flfl flfl flfl flfl flfl fl

To a stirred solution of acylfulvene (6.9 g, 32 mmol) in EtOAc (7.0 mL) was added selenium dioxide (1.75 g, 15.7 mmol) and 0.901 tert-butyl hydroperoxide (6 mL, 60 mmol). The mixture was stirred at room temperature for 24 hours and then partitioned between EtOAc and 3 x 3 mL of saturated sodium sulfite, followed by washing with saturated sodium chloride solution. The organic extracts were dried (MgSO4) and the crude product was chromatographed to give 922.5 mg (12%) of the title compound as a yellow-orange gum, and 84% yield of acylfulvene (84%).

¹ H NMR (CDCl 3): 0.76 (ddd, 1H), 1.14 (ddd, 1H), 1.27 (ddd, 1H), 1.36 (s, 3H), 1.54 (ddd, 1H), 2.03 (s, 3H) 4.02 (s, 1 H), 4.56 (s, 2 H), 6.67 (s, 1 H), 7.29 (s, 1 H).

¹³ C NMR (CDCl ₃ ): 197.9, 161.5, 146.0, 140.2, 132.9, 127.0, 119.8,

76.6, 59.9, 37.5, 27.7, 17.0, 14.7, 9.9.

MS m / e 232 (M ^& lt ^{; +} & gt ^; ), UV max wavelength (EtOH) 327.2 nm (e 7631).

Example 2

Compound 2 (a compound of formula I wherein R 1 is hydroxymethyl, R 2 is hydroxymethyl, R ₃ is hydrogen, R ₄ is methyl, R ₅ is hydroxy and R ₆ is methyl.

Add 36.3 g of paraformaldehyde to 275 ml of a 1.5 M sulfuric acid solution and 300 ml of Μβ2 <30. The mixture was stirred and heated to dissolve the solid. Then the solution was cooled to 0 ° C and 922.50 mg, 3.98 mmol of 1 was added to the solution, and the solution was stirred while allowing to warm to room temperature. After 24 hours, the orange-yellow mixture was extracted with EtOAc (2.times.300 mL), the extracts were combined, washed with sodium bicarbonate (30 mL), then brine. The organic extract was dried over magnesium sulfate. Removal of the solvent and chromatography of the residue on silica gel using ethyl acetate / hexane as eluent

4 44

4 4

4 44 «

4 4 • 4 44 444

4 4 4 4

4 4

4 4

Yield: 357.3 mg (34%) of the title product as a dark yellow-orange gum.

¹ H NMR (CDCl 3): 0-79 (ddd, 1 H), 1.16 (ddd, 1 H), 1.26 (ddd, 1 H), 1.38 (s, 3H), 1.41 (ddd, 1 H), 2.20 (s, 3H), 3.90 (s, 1 H), 4.63 (s, 2 H), 4.74 (s, 2 H), 7.25 (s, 1 H).

MS m / e 262 (M ^& lt ^{; +} & gt ^; ), UV max EtOH 330.0 nm (e 5011).

Example 3

Compound 3 (a compound of formula I wherein R f is hydrogen, R ₂ is acetoxymethyl, R ₃ is hydrogen, R ₄ is methyl, R ₅ is hydroxy and R ₆ is methyl.

To a stirred solution of Compound 1 (76.0 mg, 0.328 mmol) in 2 mL acetic anhydride was added anhydrous sodium acetate (126.2 mg, 1.54 mmol) at room temperature. The mixture was stirred at room temperature for 24 hours and then filtered to remove NaOAc. The solution was stored under reduced pressure for 4 hours, then the crude product was partitioned between EtOAc and saturated sodium bicarbonate solution, washed with saturated sodium chloride solution and dried over magnesium sulfate. After evaporation, the crude product is chromatographed on silica gel with ethyl acetate / hexane to give the title compound as a yellow gum (50 mg, 56%).

¹ H NMR (CDCl 3): 0.77 (ddd, 1H), 1.16 (ddd, 1H), 1.34 (ddd, 1H)

1H), 1.38 (s, 3H), 1.58 (ddd, 1 H), 2.06 (s, 3 H), 2.11 (s, 3 H), 4.99 (s, 2 H), 6.69 (s, 1 H), 7.27 (s, 1 H) ).

¹³ C NMR (CDCl 3): 197.9, 170.8, 163.6, 140.5, 140.0, 132.8, 126.7,

121.9, 76.7, 61.3, 37.9, 27.8, 20.9, 17.3, 15.3, 10.3.

4 4 4 4 4 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20

Example 4

Compound 4 (a compound of formula I wherein R f is acetoxymethyl, R ₂ is hydroxymethyl, R ₃ is hydrogen, R ₄ is methyl, R ₅ is hydroxy and R 8 is methyl)

To a stirred solution of compound 10 (430 mg, 1.49 mmol) in EtOAc (3 mL) was added selenium dioxide (186 mg, 1.67 mmol) followed by 0.901 tert-butyl hydroperoxide (0.40 mL, 4.0 mmol).

The mixture was stirred at room temperature for 4 days and then partitioned between EtOAc and 3 x 3 mL of saturated sodium sulfite, washed with saturated sodium chloride solution and dried over magnesium sulfate. After evaporation, the crude product is chromatographed to give 8.5 mg of the title product as a yellow gum in 2% yield.

¹ H NMR (CDCl ₃ ): 0.79 (ddd, 1 H), 1.17 (ddd, 1 H), 1.39 (s, 3H), 1.43 (ddd, 1 H), 1.52 (ddd, 1 H), 2.09 (s, 3H), 2.14 (s, 3H), 3.93 (s, 1 H), 4.65 (q, 2 H), 5.21 (q, 2 H), 7.32 (s, 1 H).

MS m / e 304 (M ⁺ ), UV max EtOH 330.6 nm (e 5950).

The intermediate of formula 10 can be prepared as follows, a. Compound 10

Anhydrous sodium acetate is added to 6 ml of the stirred solution of 1,4 g,

5.7 mmol of HMAF in acetic anhydride at room temperature. After 18 hours, the mixture was filtered and the resulting solution was stored under reduced pressure for 4 hours. The resulting material was partitioned between ethyl acetate and saturated aqueous sodium bicarbonate solution, then washed with saturated sodium chloride solution, and the resulting ethyl acetate solution was dried with magnesium sulfate. The solution was evaporated and purified by silica gel chromatography using ethyl acetate / hexane as eluent to give 1.4 g of compound 10 as a yellow-orange gum in 85% yield.

¹ H NMR (CDCl ₃ ): 0.74 (ddd, 1 H), 1.11 (ddd, 1 H), 1.36 (s, 3H), 1.51 (ddd, 1 H), 2.04 (s, 3H), 2.09 (s, 3H), 2.17 (s, 3H), 3.90 (s, 1 H), 5.10 (s, 2 H), 7.11 (s, 1 H).

Example 5

Compound 5 (a compound of formula I wherein R 1 is acetoxymethyl, R ₂ is acetoxymethyl, R ₃ is hydrogen, R ₄ is methyl, R ₅ is hydroxy and R 6 is methyl)

To a stirred solution of compound 4 (2 mg, 0.0066 mmol) in methylene chloride (15 µL) and acetic anhydride (135 µL) was added sodium acetate (1 mg, 0.038 mmol). The reaction mixture was stirred at room temperature for 4 hours, then the solid was filtered off. The mixture was stored under reduced pressure to remove acetic anhydrides. The crude product was then partitioned between ethyl acetate and saturated sodium bicarbonate solution, washed with saturated sodium chloride solution and dried over magnesium sulfate. After evaporation, the residue is chromatographed on silica gel with ethyl acetate / hexane to give 0.9 mg of the product as a yellow gum in 40% yield. ¹ H NMR (CDCl 3): 0.79 (ddd, 1 H), 1.15 (ddd, 1 H), 1.39 (s, 3H), 1.40 (ddd, 1 H), 1.58 (ddd, 1 H), 2.09 (s, 3H), 2.093 (s, 3H), 2.10 (s, 3H),

3.89 (s, 1 H), 5.08 (s, 2 H), 5.17 (s, 2 H), 7.25 (s, 1 H).

MS m / e 346 (M ⁺ ), UV max EtOH 332.1 nm (e 8378).

Example 6

Compound 6 (a compound of formula I wherein R 1 is hydroxymethyl, R ₂ is acetoxymethyl, R ₃ is hydrogen, R ₄ is methyl, R ₅ is hydroxy and R 6 is methyl)

To a stirred solution of 76.0 mg, 0.290 mmol of Compound 2 v

Sodium acetate (45.3 mg, 0.552 mmol) was added with acetic anhydride (1.5 mL). The reaction mixture was stirred at room temperature for 75 minutes and then stirred at room temperature. ·····

A ···· · · ····

4U ·· ·········· The solid is filtered off. The mixture was partitioned between ethyl acetate and saturated sodium bicarbonate solution, washed with saturated sodium chloride solution and dried over magnesium sulfate. Then the extract was evaporated under reduced pressure and the residue was chromatographed on silica gel with ethyl acetate / hexane to give 6.9 mg of compound 4 as a yellow gum, 10.0 mg of compound 2 as a yellow-orange gum, a minor amount of 3 more products including compound 5, and 10.0 mg of the title compound was obtained in 11.3% yield as a yellow gum.

¹ H NMR (CDCl 3): 0.81 (ddd, 1 H), 1.17 (ddd, 1 H), 1.38 (s, 3H), 1.60 (ddd, 1 H), 2.06 (s, 3H), 2.24 (s, 3H), 3.89 (s, 1H); 4.74 (m, 2H); 5.03 (m, 2H); 7.23 (1H).

MS m / e 304 (M ⁺ ), UV max EtOH 331.3 nm (e 5921).

Example 7

Compound of Formula 7 (a compound of Formula I wherein R 1 is hydrogen,

R ₂ is methoxymethyl, _R3 is hydrogen, _R4 is methyl, R5 is hydroxy and _R6 is methyl)

To a stirred solution of Compound 1 (300 mg, 1.29 mmol) in CH ₃ CN (10 mL ₎ was added CH ₃ I (1 mL) and Ag ₂ O (110 mg, 0.475 mmol ₎ . The mixture was stirred at room temperature for 3 days and then filtered and evaporated. The residue is chromatographed on silica gel with ethyl acetate / hexane to give 133 mg of compound 1, and 53 mg of the title product is obtained in 17% yield as a dark orange gum. ¹ H NMR (CDCl 3): 0.76 (ddd, 1 H), 1.13 (ddd, 1 H), 1.34 (ddd, 1 H), 1.38 (s, 3H), 2.05 (s, 3H), 3.41 (s, 3H), 3.96 ( s, 1H); 4.35 (s, 2H); 6.66 (s, 1H);

1 H), 7.28 (s, 1 H).

¹³ C NMR (CDCl 3): 197.8, 161.8, 143.3, 140.2, 133.1, 126.7, 120.9,

76.6, 69.6, 58.3, 37.6, 27.8, 17.1, 14.9, 10.0.

• · • ·

Example 8

Compound 8 (a compound of formula I wherein R 1 is methoxymethyl, R 2 is hydroxymethyl, R ₃ is hydrogen, R ₄ is methyl, R ₅ is hydroxy and R ₆ is methyl)

To a stirred solution of compound 11 (830 mg, 3.19 mmol) in ethyl acetate (10 mL) was added selenium dioxide (150 mg, 1.35 mmol) followed by 0.901 tert-butyl hydroperoxide (1 mL, 10.0 mmol). The reaction mixture was stirred at room temperature for 5 days and then partitioned between diethyl ether and saturated sodium sulfide, washed with saturated sodium chloride solution and dried over magnesium sulfate. After evaporation, the crude material was chromatographed to give 47.2 mg of the title compound as a dark orange gum (5%).

¹ H NMR (CDCl ₃ ): 0.74 (ddd, 1 H), 1.08 (ddd, 1 H), 1.32 (s, 3H), 1.33 (ddd, 1 H), 1.47 (ddd, 1 H), 2.08 (s, 3H), 3.14 (brs, 1 H), 3.35 (s, 3 H),

4.41 (q, 2H), 4.51 (brs, 2H), 7.28 (s, 1H).

a. Compound 11

Compound 11, used as an intermediate in this example, is a compound of formula I wherein R 1 is methoxymethyl, R 2 is methyl, R ₃ is hydrogen, R ₄ is methyl, R ₅ is hydroxy and R ₆ is methyl and can be prepared as follows.

To a stirred solution of 320 mg, 1.3 mmol of HMAF in 3 mL of Me ₃ CO was added 3 mL of methanol and 3 mL of 1 M sulfuric acid. The reaction mixture was stirred at room temperature for 24 hours and then extracted with diethyl ether. The extracts were combined, washed with saturated sodium bicarbonate and then saturated sodium chloride, and then dried over magnesium sulfate. After evaporation, the residue is chromatographed on silica gel with ethyl acetate / hexane to give 290 mg (86%) as a dark orange gum.

¹ H NMR (CDCl ₃ ): 0.62 (ddd, 1 H), 0.98 (ddd, 1 H), 1.24 (ddd, 1 H), 1.27 s, 3H), 1.37 (ddd, 1 H), 2.00 (s, 3H), 2.04 (s, 3H), 3.26 (s, 3H), 3.91 (brs, 1H), 4.29 (q, 2H), 7.0 (s, 1H).

¹³ C NMR (CDCl 3): 197.6, 159.7, 142.6, 138.8, 134.3, 129.7, 126.6,

75.9, 65.3, 57.3, 37.3, 27.3, 15.7, 13.9, 12.9, 9.1.

Example 9

Compound 9 (a compound of formula I wherein R 1 is methoxymethyl, R ₂ is methoxymethyl, R ₃ is hydrogen, R ₄ is methyl, R ₅ is hydroxy and R 6 is methyl)

To a stirred solution of Compound 8 (240 mg, 0.87 mmol) in CH ₃ CN (15 mL ₎ was added CH ₃ I (1.5 mL) and Ag ₂ O (150 mg, 0.647 mmol ₎ . The reaction mixture was stirred at room temperature for 48 h. and evaporate. The residue is chromatographed on silica gel with ethyl acetate / hexane to give the title compound as a yellow-orange gum (10 mg, 10%).

¹ H NMR (CDCl 3): 0.76 (ddd, 1 H), 1.11 (ddd, 1 H), 1.37 (s, 3H), 1.40 (ddd, 1 H), 1.53 (ddd, 1 H), 2.13 (s, 3H), 3.36 (s, 3H), 3.38 (s, 3H), 3.91 (s, 1 H), 4.37 (q, 2 H), 4.43 (q, 2 H), 7.27 (s, 1 H).

¹³ C NMR (CDCl 3): 198.0, 163.4, 142.8, 139.1, 132.6, 132.0, 127.2,

76.6, 67.2, 65.5, 58.3, 57.8, 38.1, 27.5, 16.3, 14.8, 9.8.

Example 10

Compound 12 [A compound of formula I wherein R 1 is -CH ₂ - (sulfur-bound cysteine), R ₂ is methyl, R ₃ is hydrogen, R ₄ is methyl, R 5 is hydroxy and R 6 is methyl]

An equivalent amount of cysteine was added to a solution of HMAF in acetone / 1M sulfuric acid (1: 1). The mixture was stirred at room temperature overnight. A large amount of ethyl acetate was then added and the water was removed with magnesium sulfate. It will also be added

Φ Φ 4

Φ Φ

Solid sodium bicarbonate to neutralize sulfuric acid. The solution was filtered, evaporated and the residue chromatographed to give compound 102 as a yellow gum.

¹ H NMR (CDCl 3): 0.78 (m, 1H), 0.89 (m, 1H), 1.06 (m, 1H), 1.31 (s, 3H), 1.43 (m, 1H), 2.15 (s, 3H), 2.21 (s, 3H), 2.91-4.02 (m, 8H), 7.04 (s, 1H).

Example 11

Compound 13 [A compound of formula I wherein R 1 is -CH ₂ - (N-acylcysteine, bound via a sulfur atom), R ₂ is methyl, R ₃ is hydrogen, R ₄ is methyl, R ₅ is hydroxy and R 6 is methyl]

To a solution of 36 mg, 0.146 mmol of HMAF in 3 mL of a 1: 1 mixture of 1 M sulfuric acid and acetone was added at room temperature

22.4 mg, 0.137 mmol of N-acetylcysteine.

The mixture was stirred for 22 hours and then extracted with ethyl acetate. The organic extracts were washed with saturated sodium bicarbonate and then with saturated sodium chloride, and then dried over magnesium sulfate. After evaporation, the crude product is chromatographed using ethyl acetate / hexane with 2-5% acetic acid to give 45.5 mg of compound 103 as a yellow gum in 85% yield.

¹ H NMR (CDCl 3): 0.72 (m, 1H), 1.09 (m, 1H), 1.23 (m, 1H), 1.36 (s, 3H), 1.47 (m, 1H), 2.07 (s, 3H), 2.10 (s, 3H), 2.13 (s, 3H), 2.97 (m,

1H), 3.14 (m, 1H), 3.82 (dd, 3.82), 4.80 (m, 2H), 6.56 (d, J = 7.2Hz),

7.10 (s, 1 H).

MS m / e 391 (M ⁺ ), 373, 229, 185, HRMS calcd for C ₂₀ H ₂₅ NO ₅ S 391.1455, found 391.1452.

Example 12

Compound 14 (a compound of Formula I wherein R 1 is (S) -2 - [(R) 44

-alpha-methylbenzylaminocarbonyl] -2- (acylamino) ethylthiomethyl, R ₂ is methyl, R ₃ is hydrogen, R 4 is methyl, R ₅ is hydroxy and R ₅ is methyl)

To a solution of compound 13 (40 mg, 0.102 mmol) in methylene chloride (1 mL) was added N-hydroxybenzotriazole (20 mg, 0.132 mmol), 20,,-diisopropylcarbodiimide (0.12 mmol) and 12 µL (0.12 mmol), alpha-methylbenzylamine. The mixture was stirred at room temperature for 1.5 hours and then partitioned between ethyl acetate and water. The organic extract was dried over magnesium sulfate. After evaporation, the crude product is chromatographed to give 33.6 mg of 14 as a yellow gum in 73% yield.

¹ H NMR (CDCl 3): 0.70 (m, 1H), 1.07 (m, 1H), 1.29 (m, 1H), 1.35 (s, 3H), 1.48 (d, J = 6.9Hz, 3H), 1.51 (m) (1H), 1.93 (s, 3H), 2.09 (s, 3H),

2.10 (s, 3H); 2.91 (m, 2H); 3.84 (dd, 2H); 4.61 (m, 1H); 5.03 (m, 1H); 6.64 (d, J = 7.8Hz, 1H); 7.07 (s) 1 H, 7.26 (m, 5H).

¹³ C NMR (CDCl 3): 197.5, 170.4, 169.2, 159.7, 142.6, 142.00, 138.3,

134.9, 129.8, 128.7, 127.4, 126.4, 125.9, 77.4, 52.5, 49.3, 37.7, 34.4

29.8, 27.6, 23.1, 22.1, 16.3, 14.3, 13.1, 9.5.

HRMS calcd for C28H ₃₄ N 2 O ₄ S calcd 494.2241 and 494.2225 found.

Example 13

Compound 15 (compound of Formula I wherein R 1 is (R) -2 - [(R) alpha-methylbenzylaminocarbonyl] -2- (acylamino) ethylthiomethyl, R ₂ is methyl, R ₃ is hydrogen, R ₄ is methyl, R ₅ is hydroxy and R ₆ is methyl)

Chromatography from Example 12 also yielded a total of 5.3 mg of Compound 15 as a yellow gum in 13% yield, ¹ H NMR (CDCl ₃ ): 0.70 (m, 1H), 1.07 (m, 1H), 1.32 (m, 1H), 1.34 (s, 3H), 1.45 (m, 1H), 1.48 (d, J = 6.9Hz, 3H), 2.03 (s, 3H), 2.05 (s, 6H),

2.76 (m, 1 H), 2.87 (m, 1 H), 3.73 (dd, 2 H), 4.50 (m, 1 H), 5.03 (m, 1 H), ·· · 9 ·· ··

9

9 99

6.46 (d, J = 7.5Hz, 1H), 6.77 (d, J = 7.8Hz, 1H), 7.05 (s, 1H), 7.31 (m, 5H).

HRMS calcd for C28H34N2O4S 494.2241, found 494.2238.

Example 14

Compound 16 [a compound of formula I wherein R f is -CH ₂ - (glutathione, linked via a sulfur atom), R ₂ is methyl, R ₃ is hydrogen,

R ₄ is methyl, R 5 is hydroxy and R ₆ is methyl]

Compound 16 can be prepared by the method of Example 10 except that the cysteine derivative is replaced by a glutathione derivative.

Example 15

General procedure for the preparation of compounds wherein R 1 is -CH ₂ - (sulfur-bound cysteine) -peptide, -CH ₂ - (sulfur-bound cysteine) -amino acid, -CH ₂ - (N-acylcysteine, bound via a sulfur atom) -peptide or -CH ₂ - (N-acylcysteine, linked via a sulfur atom) -amino acid.

Using solid phase peptide synthesis, the amino acids Leu, Phe, and Gly can be combined in a known manner to yield the following solid resin-bound tripeptides and dipeptides.

-Leu-Leu-Leu -Leu-Leu-Phe -Leu-Leu-Gly -Leu-Phe-Leu -Leu-Phe-Phe -Leu-Phe-Gly -Leu-Gly-Leu -Leu-Gly-Phe -Leu-Gly-Gly -Phe-Leu-Leu -Leu-Leu-Phe -Phe-Leu-Gly -Phe-Phe-Leu -Phe-Phe-Phe -Phe-Phe-Gly -Phe-Gly-Leu -Phe-Gly-Phe -Phe-Gly-Gly -Gly-Leu-Leu -Gly-Leu-Phe -Gly-Leu-Gly -Gly-Phe-Leu -Gly-Phe-Phe -Gly-Phe-Gly -Gly-Gly-Leu -Gly-Gly-Phe -Gly-Gly-Gly

-Leu-Leu -Leu-Phe -Leu-Gly -Phe-Leu -Phe-Phe -Phe-Gly -Gly-Leu -Giy-Phe -Gly-Gly

As the solid phase for these reactions, an acid resin having the following structure was used, where P is a polystyrene divinylbenzene resin.

The first N- (9-fluorenylmethoxycarbonyl) protected amino acid was coupled to the resin by forming a bond between the C-terminal end and the hydroxyl group of the resin using a mixture of DIPC (diisopropylcarbodiimide) and DMAP (4-dimethylaminopyridine). The percent recovery after binding was calculated and if less than 90%, the binding was repeated until the yield was greater than 90%. Then, the Fmoc protecting group of the first amino acid was removed by treatment with 20% piperidine in 1-methyl-2-pyrrolidinone, NMP. The Kaiser test was used to verify the outcome of the deprotection. 3 Kaiser reagents, ninhydrin, phenol and potassium cyanide were added to the sample to give a pale yellowish solution. The mixture was heated at 100 ° C for 3 minutes and if the color of the solution turned dark purple, the Fmoc group could be considered removed. Another amino group was coupled by peptide bond formation using DIPC / HOBT (1-hydroxybenzotriazole). NMP was used as the deprotection solvent, and dichloromethane was also used for washing and drying. Then the Kaiser test was used again

9

9 «

9 99

9 9

9 9 ♦ «9 9

9 9 9

9 9 9

9 99 if the color was not changed, the binding was successful. The binding reaction was repeated with additional amino acids. Compound 13 was coupled to the last amino acid or peptide using dichloromethane as solvent. NMP was not used as solvent for the coupling of compound 13 since it induced inversion of the center of chirality in cysteine and a racemic mixture was formed. In addition, HOBT was not used in the coupling reaction with Compound 13, since HOBT could also cause a mixture of products. Then, the peptide from the resin was cleaved with 10% acetic acid in dichloromethane. The products were tested in ultraviolet light and mass spectrometry. HMAF has two ultraviolet absorption maxima at 210 and 330 nm so that the presence of the products could be confirmed by the presence of the two bands. Mass spectroscopy was also used to confirm the presence of the desired products.

Using this general procedure, compound 13 was coupled to 36 of the above tripeptides and dipeptides, as well as the amino acid Leu, Phe and Gly individually, to yield 39 compounds of the invention. The N-acetyl group can be removed from these materials using conditions similar to Example 11 to give an additional 39 compounds of the invention. Thus, in the compound of formula I, R _x may preferably be Leu, Phe, Gly or any of the above tripeptides and dipeptides.

Example 16

Compound 17 (a compound of Formula I wherein R 1 is (S) -2- (2-carboxypyrrolidin-1-ylcarbonyl) propylthiomethyl, R 2 is methyl, R 3 is hydrogen, R 4 is methyl, R 5 is hydroxy and R 6 is methyl)

Compound 17 can be prepared in a manner similar to Example 10 except that cysteine is replaced by 1- (3).

94

4 4 4 4

4 4 4 9 • 4 4 4 94

4 4 4 4

44 • 4 44

mercapto-2-methyl-1-oxopropyl) -L-proline, i.e. captopril.

Example 17

Compound 18 (a compound of formula II wherein R ₂ is methyl, R ₃ is hydrogen, R ₄ is methyl, R 5 is hydroxy, R ₆ is methyl, and R 7 is hydroxymethyl.

178.5 mg of illudine S are dissolved in 2.0 ml of acetic anhydride and 96 mg of sodium acetate are added. After 24 hours, a mixture of monoacetate and diacetate is obtained. 171 mg of monoacetate were dissolved in dichloromethane and 695 mg of pyridinium dichloromethane was added. The mixture was stirred at room temperature for 21 hours. After working up the mixture, 118 mg of dehydroilludin S-monoacetate is obtained, which is dissolved in 5 ml of acetone and 3 ml of 1 M sulfuric acid, after 12 hours of standing, 66 mg of compound 18 is obtained.

¹ H NMR (CDCl 3): 0.63 (1H), 1.13 (1H), 1.23 (4H), 1.38 (4H), 2.05 (3H), 3.62 (1H), 3.76 (1H), 6.82 (1H).

Example 18

Representative pharmaceutical dosage forms comprising compounds of Formula I or II (Compound X) for medical or prophylactic use in humans will be described below.

(i) tablet 1 mg / tablet compound X 100.0 lactose 77.5 polyvinylpyrrolidone 15.0 cross-linked carmellose sodium 12.0 microcrystalline cellulose 92.5 magnesium stearate 3.0

300

99

9 9 9

9 9 9

9 9 9

9 9 9

9 9 9

• 9

9 9 99 • 9 9 9

9 9

(ii) tablet 2 mg / tablet Compound X 20.0 microcrystalline cellulose 410.0 starch 50.0 sodium starch glycolate 15.0 magnesium stearate L0 500 (iii) capsules mg / capsule Compound X 10.0 colloidal silica 1.5 lactose 465.5 pregelatinized starch 120.0 magnesium stearate 3.0 - · · · " 600.0 (iv) injection 1 (1 mg / ml) mg / ml compound X (free) 1.0 sodium hydrogen phosphate 12.0 sodium dihydrogen phosphate 0.7 sodium chloride 1.0 N sodium hydroxide, 4,5 pH adjustment to 7.0 to 7.5 as required water for injection to 1 ml

(v) injection 2 (10 mg / ml) mg / ml compound X (free) 10.0 sodium dihydrogen phosphate 0.3 sodium hydrogen phosphate 1.1 polyethylene glycol 400 200.0 0.1 N sodium hydroxide

• · · fl flfl flfl flfl fl flf fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl fl

pH adjustment to 7.0 - 7.5 as required water for injection to 1 ml (vi) aerosol mg / container Compound X 20.0 oleic acid 10.0 trichlorofluoromethane 5.000,0 dichlorodifluoromethane 10.000,0 dichlorotetrafluoroethane 5.000,0

The above types of compositions can be prepared by conventional methods.

The invention has been described in connection with various specific and preferred embodiments, but it will be apparent that many variations and modifications would be possible within the scope of the invention.

Claims (51)

PATENT CLAIMS An antitumor compound of the general formula I wherein R 1 is hydrogen, hydroxy, mercapto, amino, halogen, carboxy, nitro or - (CH 2) n - (X) - (Y), n is an integer from 0 to 4, X is -O-, thio -S-, -N (R _a ) - or absent, Y is C 3 -C 6 cycloalkyl, aryl, heteroaryl, a saccharide, amino acid, peptide residue or a 1 to 15-membered branched or unbranched hydrocarbon chain, optionally containing 1, 2 or 3 non-peroxide oxy, thio or -N (R _a ) - groups, wherein: this chain is optionally substituted on a carbon atom with 1, 2 or 3 substituents from oxo = O, hydroxy, carboxy, halogen, mercapto, nitro, -N (R b) c, C 3 -C 6 cycloalkyl, aryl, heteroaryl, saccharide residue, amino acids or peptides and at the same time the chain is saturated or unsaturated, R ₂ represents a carboxy group, C 1 -C 6 alkanoyl Φ 0 00 0 · 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 • 0 0 0 0 0 · · · 000 000 C 1 -C 6 alkoxycarbonyl, C 1 -C 6 alkyl halogen, -C (= O) NR _d R _e , a saccharide, amino acid, peptide or C 1 -C 6 alkyl residue substituted with 1 or 2 hydroxy groups, C 1 -C 6 alkoxy groups, C 1 -C 6 alkanoyloxy, carboxy, amino acid, peptide or carbohydrate residues or -C (= O) NR _d R _e , R ₃ is hydrogen, C 1 -C 6 alkyl, C 1 -C 6 alkoxy, C 1 -C 6 alkylthio, aryl, heteroaryl, aryloxy or heteroaryloxy, R ₄ is hydrogen or C 1 -C 6 alkyl, R ₅ is hydroxy, C 1 -C 6 alkoxy or C 1 -C 6 alkanoyloxy or R ₄ and R 5 together form ethylenedioxy, R ₆ is hydrogen, carboxy, C 1 -C 6 alkanoyl, C 1 -C 6 alkoxycarbonyl, halogenC 1 -C 6 alkyl, -C (= O) NR _f R _g , a saccharide, amino acid or peptide residue or C 1 -C 6 alkyl optionally substituted with 1 to 2 hydroxy substituents, C 1 -C 6 alkoxy, C 1 -C 6 alkanoyloxy, carboxy, amino acid, peptide or carbohydrate, or -C (= O) NR _f R _g , R _a represents a hydrogen atom, C 1 -C 6 alkyl, C 1 -C 6 alkanoyl, phenyl or benzyl a R b, R c, R a, R e, R f and R g independently represent a hydrogen atom, C 1 -C 6 alkyl, C 1 -C 6 alkanoyl, phenyl or benzyl or forms R _b and R _c or R _d and R _e or R f and R _g together with the nitrogen atom to which they are attached a pyrrolidine, piperidine or morpholine ring, wherein any aryl, heteroaryl , aryloxy or heteroaryloxy in the definition of Y, or R ₃ is optionally substituted with 1, 2 or 3 substituents selected from C1-C6alkyl, C1-C6alkoxy, C1 6 alkanoyl, C 1 -C C6alkanoyloxyskupina, C 1 -C 6 alkoxycarbonyl, hydroxyC1-C6alkyl, halogenC1 -C 6 alkyl, hydroxy, halogen, carboxy, mercapto, nitro or -N (Rh) (Rj), wherein R _h and R j independently represent hydrogen, C 1 -C 6 alkyl, C 1 -C 6 alkanoyl, phenyl or benzyl, or together form With a nitrogen atom to which the pyrrolidine, piperidine or morpholine ring, as well as the pharmaceutically acceptable salts thereof, are attached. 2. Antitumor agents of formula I R (I) wherein R 1 is - (CH ₂ ) n - (X) - (Y), wherein n is an integer from 0 to 4, X is oxy, thio, -N (R _a ) - or absent, Y is amino acid residue protected with 1 or 2 protecting groups, peptide residue or a 1 to 15-membered branched or unbranched hydrocarbon chain, optionally containing 1, 2 or 3 non-peroxide oxy, thio or -N (R _a ) - groups, which chain is substituted with 1 , 2 or 3 peptide residues and is saturated or unsaturated, R ₂ is hydrogen or C1-C6 alkyl, R ₃ is hydrogen, C 1 -C 6 alkyl, C 1 -C 6 alkoxy, C 1 -C C6alkylthioskupinu, aryl, heteroaryl, aryloxy or heteroaryloxy; R ₄ is hydrogen or C1 -C6 alkyl and R ₅ is hydroxy , C 1 -C 6 alkoxy or C 1 -C C6alkanoyloxyskupinu or consisting of R ₄ and R 5 together ethylenedioxy radical, R ₆ represents hydrogen, carboxyl, C 1 -C 6 alkanoyl, C 1 -C 6 alkoxycarbonyl, halogenC 1 -C 6 alkyl, -C (= O) NR _f R _g , a saccharide, amino acid or peptide residue, or C 1 -C 6 alkyl, optionally substituted with 1 to 2 substituents from the hydroxy group, • · · · 9 9 · C 1 -C 6 alkoxy, C 1 -C 6 alkanoyloxy, carboxy, amino acid, peptide or carbohydrate residues, or -C (= O) NR _f R _g , R _a is hydrogen, C 1 -C 6 alkyl, C 1 -C 6 alkanoyl, phenyl or benzyl; R b, R c, R a, R e, R f and R _g independently represent hydrogen, C 1 -C 6 alkyl, C 1 -C 6 alkanoyl, phenyl or benzyl or form R b and R _c or R _d and R _e or R f and R _g together with the nitrogen atom; to which a pyrrolidine, piperidine or morpholine ring is bonded. wherein any aryl, heteroaryl, aryloxy or heteroaryloxy group of Y or R ₃ is optionally substituted with 1, 2 or 3 substituents from C 1 -C 6 alkyl, C 1 -C 6 alkoxy, C 1 -C 6 alkanoyl, C 1 -C 6 alkanoyl, C 1 -C 6 alkoxycarbonyl, hydroxyC 1 C6alkyl, halogenC1-C6 alkyl, hydroxy, halo, carboxy, mercapto, nitro or -N (Rh) (Rj), wherein R _h and R j independently represent hydrogen, C 1 -C 6 alkyl, C 1 -C 6 alkanoyl, phenyl or benzyl or form together with the nitrogen atom to which they are attached a pyrrolidine, piperidine or morpholine ring, as well as pharmaceutically acceptable salts thereof. 3. Antitumor agents of formula I 9 9 99 99 9 9 9 9 Wherein R 1 represents a hydrogen atom, a hydroxy group, a mercapto group, an amino group, a halogen atom, a carboxy group, a nitro group or (CH ₂ ) _n - (X) ) - (Y), ⁿ is an integer from 0 to 4, X is oxy, thio, -N (R _a ) - or absent, Y is C 3 -C 6 cycloalkyl, aryl, heteroaryl, carbohydrate, amino acid or peptide residue or a 1 to 15-membered branched or unbranched carbon chain, optionally containing 1, 2 or 3 non-peroxide oxy, thio or -N (R _a ) - groups, the chain being optionally substituted with 1, 2 or 3 groups selected from oxo, hydroxy, carboxy, halogen, mercapto, nitro, -N (R _b ) (R _c ), C 3 -C 6 cycloalkyl, aryl, heteroaryl, carbohydrate, amino acid or peptide residue, the chain may be saturated or unsaturated, R ₂ is hydrogen or C 1 -C 6 alkyl, R 3 is hydrogen, C 1 - C6 alkyl, C1 6 alkoxy, C1 C6alkylthioskupinu, aryl, heteroaryl, aryloxy or heteroaryloxy; R ₄ is hydrogen or C1 -C6 alkyl, _R5 is hydroxy, C 1 -C 6 alkoxy or C 1 -C C6alkanoyloxyskupinu or _R4 and _R5 together form ethylenedioxy, R6 is carboxy, C 1 -C 6 alkanoyl, C 1 -C 6 alkoxycarbonyl, -C (= O) NR _f R _g, a saccharide residue, amino acid or peptide or a C1-C6alkyl, substituted with 1 or 2 substituents selected from C1-C6alkoxy, C 1 -C 6 alkanoyloxy, carboxy, amino acid, peptide or carbohydrate residue or -C (= O) NR _f R _g , R _a represents a hydrogen atom, C 1 -C 6 alkyl, C 1 -C 6 alkanoyl, phenyl or benzyl and R _b , R _c , Rd, R e, R f and R g independently represent hydrogen, C 1 -C 6 alkyl, C 1 -C 6 alkanoyl, phenyl or benzyl or form R _b and R _c , or R _d and R _e or R f and R _g together with the nitrogen atom to which they are attached pyrrolidine, piperidine or a morpholino ring wherein any aryl, heteroaryl, aryloxy or heteroaryloxy group Y or R 3 is optionally substituted with 1, 2 or 3 substituents from the group C 1 -C 6 alkyl, C 1 -C 6 alkoxy, C 1 -C 6 alkanoyl, 9 9 9 9 9 9 · 9 9 · 9 · ·· 99 9 9 9 ·· C 1-6 alkanoyloxy, C 1 -C 6 alkoxycarbonyl, hydroxyC 1 -C 6 alkyl, halogenC 1 -C 6 alkyl, hydroxy, halogen, carboxy, mercapto, nitro or -N (R _h ) (R j), where R _h and R j independently represent hydrogen, C 1 -C 6 alkyl, C 1 -C 6 alkanoyl, phenyl or benzyl or together with the nitrogen atom to which they are attached a pyrrolidine, piperidine or morpholine ring, as well as pharmaceutically acceptable salts thereof. Anti-cancer agents according to claims 1, 2 or 3, wherein R f represents - (CH 2) _n - (X) - (Y), n = 0 to 4, X represents an oxy group, a thio group, -N (R _a ) - or is absent and Y represents a peptide residue or C 1 -C 6 alkyl substituted with a peptide residue. Anti-cancer agents according to claim 1 or 3, wherein R f represents a hydroxy group, a halogen atom, a carboxy group, an aryl, a heteroaryl, a saccharide residue, an amino acid or a peptide. The antitumor agents of claim 1 or 3, wherein R f is hydrogen or C 1 -C 6 alkyl, optionally substituted with 1 or 2 substituents selected from hydroxy, halogen, methoxy or ethoxy. The antitumor agents of claim 1 or 3, wherein R f is - (CH ₂ ) _n - (X) - (Y), n = 1 or 2, X is oxy, thio or -N (R _a ) - and Y is C1-C6 alkyl or C2-C6 alkenyl, optionally substituted with 1 or 2 groups selected from hydroxy, carboxy, halo, mercapto, nitro, N (R b) (R _c) C 3 -C 6 cycloalkyl, aryl, heteroaryl or a radical of saccharide, amino acid, or peptide, wherein any aryl, heteroaryl, aryloxy or heteroaryloxy group of Y is optionally substituted with 1, 2 or 3 substituents from the group C 1 -C 6 alkyl, C 1 -C 6 alkoxy, C 1 -C 6 alkanoyl; 444 4 4 4 5 4 4 4 4 4 4 4 4 4 4 4 4 C 1-6 alkanoyloxy, C 1 -C 6 alkoxycarbonyl, hydroxyC 1 -C 6 alkyl, halogenC 1 -C 6 alkyl, hydroxy, halogen, carboxy, mercapto, nitro or -N (R _h ) (R j), where R _h and R j independently represent hydrogen, C 1 -C 6 alkyl, C 1 -C 6 alkanoyl, phenyl or benzyl or form together with the nitrogen atom to which they are attached a pyrrolidine, piperidine or morpholine ring, The antitumor agents of claim 1, 2 or 3, wherein R f is CH ₂ (a sulfur-bound cysteine residue) -R _x , wherein R _x is an amino acid or peptide residue of 2 to 24 amino acids. The antitumor agent of claim 8, wherein R _x is -Asp-Cys-Arg-Gly-Asp-Cys-Phe-Cys (SEQ ID NO: 4), -Asp-Gly-Arg-Cys (SEQ ID NO: 5) or -Asp-Gly-Cys-Lys-Asn-Phe-Phe-Trp-Lys-Thr-Phe-Thr-Ser-Cys (SEQ ID NO: 6). The antitumor agent according to claim 8, wherein R _x is a peptide residue containing 4 to 20 amino acid residues. Anti-cancer agents according to claim 1 or 3, wherein R 1 is -CH ₂ (a cysteine residue linked via a sulfur atom) or -CH ₂ (a Nacylcysteine residue linked via a sulfur atom). Anti-cancer agents according to claim 1, 2 or 3, wherein R-t is 2 - [(R) - alpha-methylbenzylaminocarbonyl] -2 (acylamino) ethylthiomethyl. Anti-cancer agents according to claim 1, 2 or 3, wherein R 1 is -CH ₂ - (a glutathione residue linked via a sulfur atom). • · · · · · · · · · · · · · · · ··· Anti-cancer agents according to claim 1 or 3, wherein R 1 represents a hydrogen atom, methyl, hydroxymethyl, methoxymethyl or acetoxymethyl. The antitumor agents of claim 1, wherein R ₂ is hydroxymethyl, methoxymethyl or acetoxymethyl. The antitumor agents of claim 1, wherein R ₂ is carboxy, C 1 -C 6 alkanoyl, C 1 -C 6 alkoxycarbonyl or -C (= O) NR _d R _e . The antitumor agent of claim 1, wherein R ₂ is C 1 -C 6 alkyl, substituted with 1 or 2 substituents selected from hydroxy, C 1 -C 6 alkoxy, C 1 -C 6 alkanoyloxy, carboxy, amino acid, peptide or carbohydrate, or -C (= O) NR _d R _e . The antitumor agent according to claim 1, 2 or 3, wherein R ₄ is methyl. Anti-cancer agents according to claim 1, 2 or 3, wherein R 5 is hydroxy. Anti-cancer agents according to claim 1 or 2, wherein R 6 represents a hydrogen atom. Anti-cancer agents according to claim 1, 2 or 3, wherein R 6 is carboxy, C 1 -C 6 alkanoyl, C 1 -C 6 alkoxycarbonyl or -C (= O) NR _f R _g . Anti-cancer agents according to claim 1 or 2, wherein R 6 is C 1 -C 6 alkyl, optionally substituted with 1 or 2 hydroxy groups, C 1 -C 6 alkoxy, C 1 -C 6 alkanoyloxy, carboxy, amino acid, peptide or carbohydrate, or -C (= O) NR _f R _g . The antitumor agent according to claim 1 or 2, wherein R ₆ is methyl or hydroxymethyl. The antitumor agent of claim 1, wherein R ₂ is C 1 -C 6 alkyl, substituted with hydroxy, C 1 -C 6 alkoxy, C 1 -C 6 alkanoyloxy or carboxy, R 3 is hydrogen, C 1 -C 6 alkyl, C 1 -C 6 alkoxy, C 1 -C 6 alkylthio, aryl, heteroaryl , aryloxy or heteroaryloxy, R ₄ is hydrogen or C 1 -C 6 alkyl, R 5 is hydroxy, and R ₆ is C 1 -C 6 alkyl, optionally substituted with hydroxy, C 1 -C 6 alkoxy, C 1 -C 6 alkanoyloxy or carboxy, and pharmaceutically acceptable salts thereof. Anti-cancer agents according to claim 1, wherein R 1 is hydrogen or C 1 -C 6 alkyl optionally substituted by hydroxy, halogen, methoxy, ethoxy or acetoxy, R ₂ is hydroxymethyl, methoxymethyl or acetoxymethyl, R 3 is hydrogen, R ₄ is methyl R ₅ is hydroxy and R ₅ is methyl or hydroxymethyl, as well as pharmaceutically acceptable salts thereof. The antitumor agent according to claim 2, wherein R 1 is - (CH ₂ ) _n - (X) - (Y), n = 0 to 4, X is oxy, thio, -N (R _a ) - or is absent and Y means an amino acid residue provided with one or a wood protecting group. The antitumor agent of claim 2, wherein R 1 is -CH ₂ (a sulfur-bound N-acylcysteine residue), (S) -2 - [(R) -alpha-methylbenzylaminocarbonyl] -2- (acylamino) ethylthiomethyl, or (R) -2 - [(R) -alpha-methylbenzylaminocarbonyl] -2- (9) · - (acylamino) ethylthiomethyl. A dimeric compound consisting of the two anti-tumor agents of claim 1 linked by a linker. 29th antitumor compounds of formula I wherein Ri is - (CH2) _n - (X) - (Y) n represents an integer of 1-4, preferably 1, X is oxy, thio, or -N _(Ra) - preferably a thio group, Y represents a 2 to 15-membered branched or unbranched carbon chain, respectively. containing 1, 2 or 3 non-peroxide oxy, thio or -N (R _a ) - groups, the chain is substituted on a carbon atom of 1, 2 or 3, substituents from oxo, carboxy, mercapto, -N (Rb) (R _c ) , C 3 -C 6 cycloalkyl, aryl, heteroaryl, a saccharide, amino or peptide residue and is saturated or unsaturated, preferably Y is 2 to 6, 6 membered branched or unbranched uhiíkový chain substituted on a carbon atom by 1, 2 or 3 substituents selected from oxo, heteroaryl, amino acid residue or peptide residue, R2 is hydrogen or C1C6alkyl, R ₃ is hydrogen, C1C6alkyl, C1 C 6 alkoxy, C 1 -C 6 alkylthio, aryl, heteroaryl, aryloxy or heteroaryloxy, R ₄ represents hydrogen or C 1 -C 6 alkyl and R 5 represents hydroxy, C 1 -C 6 alkoxy or C 1 -C 6 alkanoyloxy or R ₄ and R 5 together represent ethylene R6 is hydrogen, carboxy, C 1 -C 6 alkanoyl, C 1 -C 6 alkoxycarbonyl, halogenC1-C6alkyl, -C (= O) NR f R _g, a saccharide residue or a peptide or amino C1-C6 alkyl optionally substituted with 1 or 2 substituents selected from hydroxy, C 1 -C 6 alkoxy, C 1 -C 6 alkanoyloxy, carboxy, amino acid, peptide or carbohydrate residues or -C (= O) NR _f R _g , R _a represents a hydrogen atom, C 1 -C 6 alkyl, C 1 -C 6 alkanoyl, phenyl or benzyl and R b, R _c , R f and R g independently represent C 1 -C 6 alkyl, C 1 -C 6 alkanoyl, phenyl or benzyl or form R b and R _c or R f and R g together with the nitrogen atom to which they are attached a pyrrolidine, piperidine or morpholine ring, wherein any aryl, heteroaryl, aryloxy or heteroaryloxy group Y or R ₃ is optionally substituted with 1, 2 or 3 substituents from the group C 1 -C 6 alkyl (, C 1 -C 6 alkoxy, C 1 -C 6 alkanoyl, C 1-6 alkanoyloxy, C1-C6alkoxycarbonyl, hydroxyC1-C6alkyl, halogenC1-C6alkyl, hydroxy, halogen, carboxy, mercapto, nitro or -N (Rh) (Rj), wherein Rh and Rj independently represent hydrogen, C1-C6alkyl, C1-C6alkanoyl, phenyl or benzyl or together with the nitrogen atom to which they are attached a pyrrolidine, piperidine or morpholine ring, as well as pharmaceutically acceptable salts thereof. The antitumor agent of claim 29, which is or a pharmaceutically acceptable salt thereof. • ······· · · · · · ··· ··· ·· ·· 31. An antitumor compound of formula II _R2 is C1-C6alkyl, R ₃ is hydrogen, C 1 -C 6 alkyl, C 1 -C 6 alkoxy, C 1 -C 6 alkylthio, aryl, heteroaryl, aryloxy or heteroaryloxy, R ₄ represents a hydrogen atom or C 1 -C 6 alkyl and R ₅ is hydroxy, C 1 -C 6 alkoxy or C 1 -C 6 alkanoyloxy or R ₄ and R ₅ together are ethylenedioxy, R 6 is hydrogen, carboxy, C 1 -C 6 alkanoyl, C 1 -C 6 alkoxycarbonyl, halogenC 1 -C 6 alkyl, -C (= O) NR _f R _g , a carbohydrate, amino acid or peptide residue or C 1 -C 6 alkyl optionally substituted with 1 or 2 substituents from hydroxy, C 1 -C 6 alkoxy, C 1 -C 6 alkanoyloxy, carboxy, amino acid, peptide or carbohydrate, or -C (= O) NR _f R g, R 2 represents a carboxy group, C 1 -C 6 alkanoyl, C 1 -C 6 alkoxycarbonyl, halogenC 1 -C 6 alkyl, -C (= O) NR _d R _e , a saccharide, amino acid or peptide residue or C 1 -C 6 alkyl, optionally substituted with 1 or 2 substituents from hydroxy, C 1 -C 6 C 6 alkoxy, C 1 -C 6 alkanoyloxy, carboxy, amino acid, peptide or carbohydrate residue, or -C (= O) NR d R _e , · · · · · · · · · · · · · · · · · · · · · · · · · ·· R _d , R _e , R f and R _g independently represent hydrogen, C 1 -C 6 alkyl, C 1 -C 6 alkanoyl, phenyl or benzyl or form R _d and R _e or R f and R _g together with the nitrogen atom to which they are attached pyrrolidine, piperidine or a morpholine ring, wherein any aryl, heteroaryl, aryloxy or heteroaryloxy group R ₃ is optionally substituted with 1, 2 or 3 substituents from C 1 -C 6 alkyl, C 1 -C 6 alkoxy, C 1 -C 6 alkanoyl, C 1-6 alkanoyloxy, C 1 -C 6 alkoxycarbonyl, hydroxyC 1 , halogenC1-C6alkyl, hydroxy, halogen, carboxy, mercapto, nitro or -N (Rh) (Rj), wherein Rh and Rj independently represent hydrogen, C1-C6alkyl, C1-C6alkanoyl, phenyl or benzyl or form together with the atom nitrogen to which a pyrrolidine, piperidine or morpholine ring is bound, as well as pharmaceutically acceptable salts thereof. The antitumor agent of claim 32, wherein R 7 is C 1 -C 6 alkyl, substituted with 1 or 2 substituents selected from hydroxy, C 1 -C 6 alkoxy, C 1 -C 6 alkanoyloxy, carboxy, amino acid, peptide or carbohydrate, or -C (= O) NR _d R _e . The antitumor agent of claim 32, wherein R 7 is hydroxymethyl. The antitumor agent of claim 32, which is or a pharmaceutically acceptable salt thereof. φ φ An antitumor agent comprising a compound according to claim 1, 2, 3, 29 or 32 and a reagent capable of binding to a tumor-associated antigen. 36. The antitumor agent of the general formula wherein A is - (CH _{2) n} - (X) - (Y) n = 1-4, X is oxy, thio, or -N (R _a ) -, Y is a peptide residue or C 1 -C 6 alkyl substituted with a peptide residue, _R2 is C1-C6alkyl, R ₃ is hydrogen, C 1 -C 6 alkyl, C 1 -C 6 alkoxy, C 1 -C 6 alkylthio, aryl, heteroaryl, aryloxy or heteroaryloxy, R ₄ is hydrogen or C 1 -C 6 alkyl; R 5 is hydroxy, C 1 -C 6 alkoxy or C 1 -C 6 alkanoyloxy or R ₄ and R ₅ together form ethylenedioxy, R ₆ represents hydrogen, carboxyl, C 1 -C 6 alkanoyl, ♦ 9 9 · «· 9 9 99 99 • ♦ ♦ 9 9 9 9 9 • 9 9 9 9 C 1 -C 6 alkoxycarbonyl, halogenC 1 -C 6 alkyl, -C (= O) NR _f R _g , a carbohydrate, amino acid or peptide residue or C 1 -C 6 alkyl, optionally substituted with 1 or 2 substituents from hydroxy, C 1 -C 6 alkoxy, C 1 -C 6 alkanoyloxy, carboxy, amino acid , peptides or carbohydrate or the group -C (= O) NR _f R _g , Rd, R _e , R f and R _g independently represent hydrogen, C 1 -C 6 alkyl, C 1 -C 6 alkanoyl, phenyl or benzyl or form Rd and R _e or R f and R _g together with the nitrogen atom to which they are attached pyrrolidine, piperidine or morpholine ring, wherein any aryl, heteroaryl, aryloxy or heteroaryloxy group R ₃ is optionally substituted with 1, 2 or 3 substituents from C 1 -C 6 alkyl, C 1 -C 6 alkoxy, C 1 -C 6 alkanoyl, C 1-6 alkanoyloxy, C 1 -C 6 alkoxycarbonyl, hydroxyC 1 -C 6 alkyl, halogenC 1 , hydroxy, halogen, carboxy, mercapto, nitro or -N (Rh) (Rj), wherein Rh and Rj are independently hydrogen, C 1 -C 6 alkyl, C 1 -C 6 alkanoyl, phenyl or benzyl or form together with the nitrogen atom to which they are attached are bound by a pyrrolidine, piperidine or morpholine ring, as well as pharmaceutically acceptable salts thereof. 37. Antitumor agents of formula I wherein Ri means - (ΟΗ ₂ ) _Π - (Χ) - (Υ) n = 1 to 4, ·· 00 • 0 ·· ·· • · 0 0 0 0 0 · · 0 0 · 00 00 X is oxy, thio, or -N (R _a ) -, Y is C 2 -C 6 alkenyl, optionally substituted with 1 or 2 substituents from oxo, hydroxy, carboxy, halogen, mercapto, nitro, -N (R _b ) (R _c ), C 3 -C 6 cycloalkyl, aryl, heteroaryl, saccharide residue, amino acids or a peptide, _R2 is C1-C6alkyl, R ₃ is hydrogen, C 1 -C 6 alkyl, C 1 -C 6 alkoxy, C 1 -C C6alkylthioskupinu, aryl, heteroaryl, aryloxy or heteroaryloxy, R ₄ represents a hydrogen atom or C 1 -C 6 alkyl and R 5 is hydroxy, C 1 -C 6 alkoxy or C 1 -C 6 alkanoyloxy or R ₄ and R ₅ together form ethylenedioxy, R ₆ is hydrogen, carboxy, C 1 -C 6 alkanoyl, C 1 -C 6 alkoxycarbonyl, haloC 1 -C 6 alkyl, -C (= O) NR _f R _g , a saccharide, amino acid or peptide residue or C 1 -C 6 alkyl optionally substituted with 1 or 2 substituents from the group hydroxy, C1-C6alkoxy, C1-C6alkanoyloxyskupina, carboxy, amino acid residue, peptide or carbohydrate or -C (= O) NR _f R _g, R _d , R _e , R f and R _g independently represent hydrogen, C 1 -C 6 alkyl, C 1 -C 6 alkanoyl, phenyl or benzyl or form R _d and R _e or R f and R _g together with the nitrogen atom to which they are attached pyrrolidine, piperidine or a morpholine ring, wherein any aryl, heteroaryl, aryloxy or heteroaryloxy group R ₃ is optionally substituted with 1, 2 or 3 substituents from C 1 -C 6 alkyl, C 1 -C 6 alkoxy, C 1 -C 6 alkanoyl, C 1-6 alkanoyloxy, C 1 -C 6 alkoxycarbonyl, hydroxyC 1 , halogenC 1 -C 6 alkyl, hydroxy, halogen, carboxy, mercapto, nitro or. 4 · · · ·· ·· -N (Rh) (R j), wherein R _h and R j independently represent hydrogen, C 1 -C 6 alkyl, C 1 -C 6 alkanoyl, phenyl or benzyl or form together with the nitrogen atom to which they are attached a pyrrolidine, piperidine or morpholine ring, as well as pharmaceutically acceptable salts thereof. 38. The antitumor agent according to claim 1, wherein: R 1 is -CH 2 - (a sulfur-bound cysteine residue) -R _x , where R _x is an amino acid or peptide residue of 2 to 24 amino acids, _R2 is C1-C6alkyl, R ₃ is hydrogen, C 1 -C 6 alkyl, C 1 -C 6 alkoxy, C 1 -C C6alkylthioskupinu, aryl, heteroaryl, aryloxy or heteroaryloxy, R ₄ represents a hydrogen atom or C 1 -C 6 alkyl and R ₅ is hydroxy, C 1 -C 6 alkoxy or C 1 -C 6 alkanoyloxy or R ₄ and R ₅ together form ethylenedioxy, R ₆ is hydrogen, carboxy, C 1 -C 6 alkanoyl, C 1 -C 6 alkoxycarbonyl, haloC 1 -C 6 alkyl, -C (= O) NR _f R _g , a carbohydrate, amino acid or peptide residue or C 1 -C 6 alkyl optionally substituted with 1 or 2 hydroxy substituents, C 1 -C 6 alkoxy, C 1 -C 6 alkanoy (oxy, carboxy, amino acid, peptide or carbohydrate residue, or -C (= O) NR _f R _g , R _d , R _e , R f and R _g independently represent hydrogen, C 1 -C 6 alkyl, 00 00 0 0 00 00 • · · 0 0 0 · 0 0 0 Φ • 0 0 0 00 00 0 0 0 0 0 0 0 0 0 0 0 0 0 0 00 C 1 -C 6 alkanoyl, phenyl or benzyl or forms R _d and R _e or R f and R _g together with the nitrogen atom to which they are attached a pyrrolidine, piperidine or morpholine ring, wherein any aryl, heteroaryl, aryloxy or heteroaryloxy group R ₃ is optionally substituted 1, 2 or 3 substituents from the group C1-C6alkyl, C1-C6alkoxy, C1-C6alkanoyl, C1-6alkanoyloxy, C1-C6alkoxycarbonyl, hydroxyC1-C6alkyl, halogenC1-C6alkyl, hydroxy, halogen, carboxy, mercapto, nitro or -N (-) R h) (R j) wherein R h and R j independently represent hydrogen, C 1 -C 6 alkyl, C 1 -C 6 alkanoyl, phenyl or benzyl or form together with the nitrogen atom to which they are attached a pyrrolidine, piperidine or morpholine ring, as well as pharmaceutically acceptable salts thereof substances. A compound according to claim 1, wherein R 1 is -CH ₂ - (a sulfur-bound N-acylcysteine residue) R _x , wherein R _x is an amino acid or peptide residue of 2 to 24 amino acids, R ₂ represents C 1 -C 6 alkyl, R ₃ represents hydrogen, C 1 -C 6 alkyl, C 1 -C 6 alkoxy, C 1 -C C6aikylthioskupinu, aryl, heteroaryl, aryloxy or heteroaryloxy, R ₄ represents a hydrogen atom or C 1 -C 6 alkyl and R ₅ is hydroxy, C 1 -C 6 alkoxy or 9 • 9 " 9 9 9,999 9,9 9 9 9 9 9 9 9 9 9 C 1 -C 6 alkanoyloxy or R ₄ and R ₅ together form ethylenedioxy, R ₆ is hydrogen, carboxy, C 1 -C 6 alkanoyl, C 1 -C 6 alkoxycarbonyl, halogenC 1 -C 6 alkyl, -C (= O) NR _f R _g , a carbohydrate, amino acid or peptide residue, or C 1 -C 6 alkyl, optionally substituted with 1 or 2 substituents from hydroxy, C1-C6alkoxy, C1-C6alkanoyloxy, carboxy, *. an amino acid residue, a peptide or a carbohydrate, or a group -C (= O) NR _f R _g , Rd, R e, R f and R g independently represent hydrogen, C 1 -C 6 alkyl, C 1 -C 6 alkanoyl, phenyl or benzyl or when R <I R _e or R f and R _g together with the nitrogen atom to which they are attached form a pyrrolidine, piperidine or morpholine ring wherein any aryl, heteroaryl, aryloxy or heteroaryloxy R ₃ is optionally substituted 1, 2 or 3 C 1 -C 6 alkyl, C 1 -C 6 alkoxy substituents, C1-C6alkanoyl, C1-6alkanoyloxy, C1-C6alkoxycarbonyl, hydroxyC1-C6alkyl, halogenC1-C6alkyl, hydroxy, halogen, carboxy, mercapto, nitro or -N (Rh) (Rj), wherein Rh and Rj independently represent hydrogen, C1 -C6alkyl, C1-C6alkanoyl, phenyl or benzyl, or together with the nitrogen atom to which they are attached a pyrrolidine, piperidine or morpholine ring, as well as pharmaceutically acceptable salts thereof. Anti-cancer agents according to claim 1, of the general formula I (I) 9 9 9 9 9 9 99 99 where 9 R 1 is -CH 2 - (a glutathione residue bonded via a sulfur atom), R2 is C1-C6alkyl, R ₃ is hydrogen, C 1 -C 6 alkyl, C 1 -C 6 alkoxy, C 1 -C 6 alkylthio, aryl, heteroaryl, aryloxy or heteroaryloxy, R ₄ represents a hydrogen atom or C 1 -C 6 alkyl and R 5 is hydroxy, C 1 -C 6 alkoxy or C 1 -C 6 alkanoyloxy or R ₄ and R ₅ together form ethylenedioxy, R ₆ is hydrogen, carboxy, C 1 -C 6 alkanoyl, C 1 -C 6 alkoxycarbonyl, halogenC 1 -C 6 alkyl, -C (= O) NR _f R _g , a carbohydrate, amino acid or peptide residue or C 1 -C 6 alkyl optionally substituted with 1 or 2 hydroxy substituents, C 1 -C 6 alkoxy, C 1 -C 6 alkanoyloxy, carboxy, amino acid, peptide or carbohydrate residues, or -C (= O) NR _f R _g , R _d , R _e , R f and R _g independently represent hydrogen, C 1 -C 6 alkyl, C 1 -C 6 alkanoyl, phenyl or benzyl or form R _d and R _e or R f and R _g together with the nitrogen atom to which they are attached pyrrolidine, piperidine or a morpholine ring, wherein any aryl, heteroaryl, aryloxy or heteroaryloxy group R ₃ is optionally substituted with 1,2 or 3 substituents from C 1 -C 6 alkyl, C 1 -C 6 alkoxy, C 1 -C 6 alkanoyl, C 1-6 alkanoyloxy, C 1 -C 6 alkoxycarbonyl, hydroxyC 1 -C 6 , halogenC1-C6alkyl, hydroxy, halogen, carboxy, mercapto, nitro or -N (Rh) (Rj), wherein Rh and Rj independently represent hydrogen, C1-C6alkyl, C1-C6alkanoyl, phenyl or benzyl or form together with the atom nitrogen to which a pyrrolidine, piperidine or morpholine ring is bound, as well as pharmaceutically acceptable salts thereof. 9 · 99 ti • 9 · 9 9 99 9 9 9 9 99 ·· 99 99 9 9 9 99 99 41. The antitumor agent according to claim 1, wherein: EMI15.0 [0100] Rt is 2 - [(R) -alpha-methylbenzylaminocarbonyl] -2 (acylamino) ethylthiomethyl, _R2 is C1-C6alkyl, R ₃ is hydrogen, C 1 -C 6 alkyl, C 1 -C 6 alkoxy, C 1 -C C6alkylthioskupinu, aryl, heteroaryl, aryloxy or heteroaryloxy, R ₄ represents a hydrogen atom or C 1 -C 6 alkyl and R ₅ is hydroxy, C 1 -C 6 alkoxy or C 1 -C 6 alkanoyloxy or R ₄ and R ₅ together are ethylenedioxy, R ₆ is hydrogen, carboxy, C 1 -C 6 alkanoyl, C 1 -C 6 alkoxycarbonyl, halogenC 1 -C 6 alkyl, -C (= O) NR _f R _g , a carbohydrate, amino acid or peptide residue, or C 1 -C 6 alkyl optionally substituted with 1 or 2 hydroxy substituents, C 1 -C 6 alkoxy, C 1 -C 6 alkanoyloxy, carboxy, amino acid, peptide or carbohydrate, or -C (= O) NR _f R _g , Rd, R _e , R f and R _g independently represent hydrogen, C 1 -C 6 alkyl, C 1 -C 6 alkanoyl, phenyl or benzyl or form R _d and R _e or R f and R _g together with the nitrogen atom to which they are attached pyrrolidine, piperidine or morpholine ring , φ · · · φ φ φ φ φ φ φ φ φ φ · · φ φ φ φ · · · · · φ φ φ φ φ φ φ φ φ φ lová lová lová lová group, aryloxy or heteroaryloxy R ₃ is optionally substituted with 1, 2 or 3 substituents selected from C1-C6alkyl, C1-C6alkoxy, C1 6 alkanoyl, C1-6alkanoyloxyskupina, C 1 -C 6 alkoxycarbonyl, hydroxyC1-C6alkyl, halogenC1-C6 alkyl, hydroxy, halogen , carboxy, mercapto, nitro or -N (Rh) (Rj), wherein Rh and Rj independently represent hydrogen, C1-C6alkyl, C1-C6alkanoyl, phenyl or benzyl or form together with the nitrogen atom to which they are attached a pyrrolidine, piperidine or a morpholine ring; pharmaceutically acceptable salts thereof. 42. The antitumor agent of claim 1, wherein R 1 is - (CH ₂ ) _n - (X) - (Y), n = 1 to 4, X is oxy, thio, or -N (R _a ) -, Y represents a branched or unbranched carbon chain of 2 to 15 carbon atoms, substituted on the carbon atom by 1 or 2 substituents from the group carboxy, C 3 -C 6 cycloalkyl, heteroaryl, a sugar, amino acid or peptide residue, said chain being saturated or unsaturated, _R2 is C1-C6alkyl, R ₃ is hydrogen, C 1 -C 6 alkyl, C 1 -C 6 alkoxy, C 1 -C C6alkylthioskupinu, aryl, heteroaryl, aryloxy or heteroaryloxy, R ₄ represents a hydrogen atom or C 1 -C 6 alkyl and R 5 is hydroxy, C 1 -C 6 alkoxy or C 1 -C 6 alkanoyloxy or R ₄ and R ₅ together form ethylenedioxy, R ₆ is hydrogen, carboxy, C 1 -C 6 alkanoyl, C 1 -C 6 alkoxycarbonyl, haloC 1 -C 6 alkyl, -C (= O) NR _f R _g , a carbohydrate, amino acid or peptide residue or C 1 -C 6 alkyl optionally substituted with 1 or 2 hydroxy substituents, C 1 -C 6 alkoxy, C 1 -C 6 alkanoyloxy, carboxy, amino acid, peptide or carbohydrate, or -C (= O) NR _f R _g , Rd, R _e , R f and R _g independently represent hydrogen, C 1 -C 6 alkyl, C 1 -C 6 alkanoyl, phenyl or benzyl or form R _d and R _e or R f and R _g together with the nitrogen atom to which they are attached pyrrolidine, piperidine or morpholine a ring, wherein any aryl, heteroaryl, aryloxy or heteroaryloxy group R ₃ is optionally substituted with 1, 2 or 3 substituents from C 1 -C 6 alkyl, C 1 -C 6 alkoxy, C 1 -C 6 alkanoyl, C 1-6 alkanoyloxy, C 1 -C 6 alkoxycarbonyl, hydroxyC 1 -C 6 alkyl, halogenC 1 -C 6 alkyl -C6alkyl, hydroxy, halogen, carboxy, mercapto, nitro or -N (Rh) (Rj), wherein Rh and Rj independently represent hydrogen, C1-C6alkyl, C1-C6alkanoyl, phenyl or benzyl or form together with the nitrogen atom, to which are bound a pyrrolidine, piperidine or morpholine ring, as well as pharmaceutically acceptable salts thereof. The antitumor agents according to claim 1, of the general formula (I), wherein the flfl flfl flfl flfl. R 1 is - (CH 2) n - (X) - (O. n = 1, X is thio, Y represents a 2 to 6-membered branched or unbranched carbon chain substituted on the carbon atom by 1, 2 or 3 substituents selected from oxo, heteroaryl or the remainder of the peptide, _R2 is C1-C6alkyl, R ₃ is hydrogen, C 1 -C 6 alkyl, C 1 -C 6 alkoxy, C 1 -C C6alkylthioskupinu, aryl, heteroaryl, aryloxy or heteroaryloxy, R ₄ represents a hydrogen atom or C 1 -C 6 alkyl and R 5 is hydroxy, C 1 -C 6 alkoxy or C 1 -C 6 alkanoyloxy or R ₄ and R ₅ together form ethylenedioxy, R 6 is hydrogen, carboxy, C 1 -C 6 alkanoyl, C 1 -C 6 alkoxycarbonyl, halogenC 1 -C 6 alkyl, -C (= O) NR _f R _g , a carbohydrate, amino acid or peptide residue or C 1 -C 6 alkyl optionally substituted with 1 or 2 substituents from hydroxy, C 1 -C6alkoxy, C1-C6alkanoyloxy, carboxy, amino acid, peptide or carbohydrate, or -C (= O) NR _f R _g , R _d , R _e , R f and R _g independently represent hydrogen, C 1 -C 6 alkyl, C 1 -C 6 alkanoyl, phenyl or benzyl or form R _d and R _e or R f and R _g together with the nitrogen atom to which they are attached pyrrolidine, piperidine or a morpholine ring, wherein any aryl, heteroaryl, aryloxy or heteroaryloxy group R 3 is optionally substituted with 1, 2 or 3 substituents from C 1 -C 6 alkyl, C 1 -C 6 alkoxy, C 1 -C 6 alkanoyl, C 1-6 alkanoyloxy, C 1 -C 6 alkoxycarbonyl, hydroxyC 1 -C 6 haloC 1 -C 6 alkyl, hydroxy, halogen, carboxy, mercapto, nitro or -N (Rh) (Rj) wherein Rh and Rj independently represent hydrogen, C1-C6alkyl, C1-C6alkanoyl, phenyl or benzyl or form together with the nitrogen atom to which they are attached a pyrrolidine, piperidine or morpholine ring, as well as a pharmaceutically acceptable acceptable salts thereof. 44. The antitumor agent according to claim 1, wherein the compound of formula I is a compound of formula (I) R 1 is hydroxy, carboxy, aryl, heteroaryl or a saccharide, amino acid or peptide residue, _R2 is C1-C6alkyl, R ₃ is hydrogen, C 1 -C 6 alkyl, C 1 -C 6 alkoxy, C 1 -C C6alkylthioskupinu, aryl, heteroaryl, aryloxy or heteroaryloxy, R ₄ represents a hydrogen atom or C 1 -C 6 alkyl and R ₅ is hydroxy, C 1 -C 6 alkoxy or C 1 -C 6 alkanoyloxy or R ₄ and R 5 together are ethylenedioxy, R ₆ is hydrogen, carboxy, C 1 -C 6 alkanoyl, C 1 -C 6 alkoxycarbonyl, haloC 1 -C 6 alkyl, -C (= O) NR _f R _g , a saccharide, amino acid or peptide residue or C 1 -C 6 alkyl optionally substituted with 1 or 2 hydroxy substituents, C 1 -C 6 alkoxy, C 1 -C 6 alkanoyloxy, carboxy, amino acid, peptide or carbohydrate, or -C (= O) NR _f R _g , Rd, R e, R f and R _g independently represent hydrogen, C 1 -C 6 alkyl, C 1 -C 6 alkanoyl, phenyl or benzyl or form R _d and R _e or R f and R _g together with the nitrogen atom to which they are attached pyrrolidine, piperidine or morpholine a ring, wherein any aryl, heteroaryl, aryloxy or heteroaryloxy R ₃ is optionally substituted with 1, 2 or 3 substituents from C 1 -C 6 alkyl, C 1 -C 6 alkoxy, C 1 -C 6 alkanoyl, C 1-6 alkanoyloxy, C 1 -C 6 alkoxycarbonyl, hydroxyC 1 -C 6 alkyl, C 6 alkyl, hydroxy, halogen, carboxy, mercapto, nitro or -N (Rh) (R j), wherein R _h and R j independently represent hydrogen, C 1 -C 6 alkyl, C 1 -C 6 alkanoyl, phenyl or benzyl or form together with the nitrogen atom, to which a pyrrolidine, piperidine or morpholine ring, as well as pharmaceutically acceptable salts thereof, are bonded. 45. A pharmaceutical composition comprising as an active ingredient an antitumor agent according to any one of claims 1, 2, 3, 29, 30, 31, 34, 36, 37, 38, 39, 40; 41, 42, 43 or 44 in combination with a pharmaceutically acceptable diluent or carrier. The antitumor agents according to claims 1, 2, 3, 29, 30, 31, 34, 36, 37, 38, 39, 40, 41, 42, 43 or 44 for use in medical purposes. The antitumor agent according to claim 46 for use in the treatment of cancer. Use of the antitumor agents according to claims 1, 2, 3, 29, 30, 31 34, 36, 37, 38, 39, 40, 41, 42, 43 or 44 for the manufacture of a pharmaceutical composition for the treatment of cancer. The use of claim 48, wherein the cancer is a solid tumor. • 9 50. A method of inhibiting tumor cells, comprising contacting the cells in vitro or in vivo with an effective amount of the antitumor agents of any one of claims 1, 2, 3, 29, 30, 50, 50, 50, 31, 34, 36, 37, 38, 39, 40, 41.42, 43 or 44. 51. A method of treating cancer comprising administering an effective amount of an antitumor agent according to any one of claims 1, 2, 3, 29, 30, 31, 34, 36, 37, 38, 39, 40, 41. 42, 43 or 44.