HK1184363B - Novel auristatin derivatives and use thereof - Google Patents

Abstract

The invention relates to novel derivatives of monomethyl auristatin F, to methods for producing said derivatives, to the use of said derivatives to treat and/or prevent illnesses, and to the use of said derivatives to produce drugs for treating and/or preventing illnesses, in particular hyperproliferative and/or angiogenic diseases such as cancer diseases. Such treatments can occur as monotherapy or in combination with other drugs or further therapeutic measures.

Description

This notification concerns novel derivatives of monomethylauristatine F, the methods of manufacture of these derivatives, the use of these derivatives for the treatment and/or prevention of diseases and the use of these derivatives for the manufacture of medicinal products for the treatment and/or prevention of diseases, in particular hyperproliferative and/or angiogenic diseases such as cancer, which may be administered as monotherapy or in combination with other medicinal products or other therapeutic measures.

Cancer is the result of uncontrolled cell growth in a variety of tissues. In many cases, the new cells enter existing tissues (invasive growth), or they metastasize to distant organs. Cancer occurs in a variety of organs and often has tissue-specific disease processes. Therefore, the term cancer describes a large group of defined diseases of different organs, tissues, and cell types.

Early-stage tumours can be removed by surgery and radiotherapy, if necessary.Metastatic tumours can usually only be treated by chemotherapy as a palliative, with the aim of achieving the optimal combination of improving quality of life and prolonging life.

Most chemotherapeutic agents currently administered parenterally are often not targeted at tumour tissue or tumour cells but are distributed non-specifically in the body by systemic administration, including to sites where exposure is undesirable, such as healthy cells, tissues and organs.

The improved and selective availability of these chemotherapeutic agents in the tumour cell or the tissue immediately surrounding it, and the associated intensification of action on the one hand and minimisation of toxic side effects on the other, have therefore been the focus of the development of new chemotherapeutic agents for several years.

Err1:Expecting ',' delimiter: line 1 column 864 (char 863)

Err1:Expecting ',' delimiter: line 1 column 184 (char 183)

Auristatin E (AE) and monomethylauristatin E (MMAE) are synthetic analogues of dolastatins, a special group of linear pseudopeptides originally isolated from marine sources, some of which exhibit very potent cytotoxic activity against tumour cells (for an overview see, for example, G. R. Pettit, Prog. Chem. Org. Nat. Prod. 70, 1-79 (1997); G. R. Pettit et al., Anti-Cancer Drug Design 10, 529-544 (1995); G. R. Pettit et al., Anti-Cancer Drug Design 13, 243-277 (1998)).

However, MMAE has the disadvantage of a relatively high systemic toxicity and, when used as antibody-agent conjugates (immunoconjugates), is not compatible with antibody-agent binding units (linkers) that do not have enzymatically cleavage target sites (S.O. Doronina et al., Bioconjugates Chem. 17, 114-124 (2006)).

Monomethylauristatin F (MMAF) is an auristatin derivative with a C-terminal phenylalanine unit that has only moderate anti-proliferative effects compared to MMAE. This is very likely due to the free carboxyl group, which due to its polarity and charge negatively affects the cell viability of this compound. In this context, the methyl ester of MMAF (MMAF-OMe) has been described as a neutral charged, cellular prodrug derivative that shows a multi-order increase in in vitro cytotoxicity compared to MMAF in several cancer cell lines [S. O. Doronina et al., 17-114-124 (2006) ]. It is assumed that this effect is produced by the absorption of MMAF by the cell, which is rapidly released by the intrinsically hydrogenous ester of MMAF. Other

However, active substance compounds based on simple ester derivatives are generally subject to the risk of chemical instability due to non-specific ester hydrolysis, independent of the intended action, for example by esterases present in blood plasma, which may significantly limit the applicability of such compounds in therapy.

The present invention was therefore intended to identify new compounds based on the only moderately active monomethylauristatin F (MMAF) and to provide for the treatment of cancer in particular, which have significantly higher cytotoxic activity in whole cell assays and higher plasma stability compared to simple ester derivatives such as MMAF-OMEs.

Monomethylauristatin F (MMAF) and various ester and amide derivatives thereof were disclosed in WO 2005/081711-A2. Other auristatin analogues with a C-terminal amide-substituted phenylalanine unit are described in WO 01/18032-A2.

The present invention is now directed to compounds of general formula (I) Other in which R1 means hydrogen, (C1-C6) alkyl or a group of the formula Q1-L1-* or Q2-L2-* wherein Other * marks the point of attachment with the nitrogen atom, Q1 hydroxycarbonyl, (C1-C4) alkoxycarbonyl or benzyloxycarbonyl means L1 means straight chain (C1-C12) alkandiyl that can be substituted up to four times with methyl and in which (a) two carbon atoms in a 1,2, 1,3, or 1,4-relation to each other can be broken into a (C3-C6) cycloalkyl or a phenyl ring, including any carbon atoms between them, or (b) up to three non-adjacent CH2 groups can be replaced by -O- Q2 hydroxy, amino or mono- ((C1-C4) -alkylamino means and Other L2 straight chain (C2-C12) alkandiyl means that can be substituted up to four times with methyl and in which (a) two carbon atoms in 1,(b) up to three non-adjacent CH2 groups can be replaced by -O-,R2 stands for methyl or hydroxy,R3 stands for hydrogen or methyl,R4 isopropyl, isobutyl, sec-butyl, tert-butyl, phenyl, benzyl, 4-hydroxybenzyl, 4-hydroxy-3-benzyl, 1-phenylethyl, diphenylmethyl, 1-H-imidazole-methyl or 1-H-indol-methyl, or R3 and R4 together with the carbon atom to which they are both bound, form a 2-phenylcyclopropane-1,1-diyl group with the formula Other Other, including: Other # marks the junctions with the rest of the molecule, and the ring A with the N-O grouping contained therein for a monocyclic or bicyclic, where appropriate substituted heterocycle of the formula Other It says in **the linking point is marked with the carbonyl group,Other R5 and R6 mean hydrogen, hydroxy, (C1-C4) -alkoxy or benzyloxy respectively, andR7 means hydrogen, fluorine, chlorine, cyano, methyl or trifluoromethyl, and their salts, solvates and solvates of salts.

The compounds of the invention are the compounds of formula (I) and their salts, solvates and solvates of salts, the compounds of formula (I) and their salts, solvates and solvates of salts, and the compounds of formula (I) and their salts, solvates and solvates of salts, as exemplified below, in so far as the compounds of formula (I) and their salts, solvates and solvates of salts are not already salts, solvates and solvates of salts.

The compounds of the invention may exist in different stereoisomeric forms, i.e. as configuration isomers or, where appropriate, as conformational isomers (enantiomers and/or diastereomers, including those in atropisomers), depending on their structure. The present invention therefore covers the enantiomers and diastereomers and their respective mixtures. From such mixtures of enantiomers and/or diastereomers, the stereoisomeric unitary components can be isolated in a known manner, using chromatographic methods, in particular HPLC chromatography at achiral and chiral phase.

If the compounds of the invention can be found in more than one tautomeric form, the present invention covers all tautomeric forms.

The salts of the present invention are those which are physiologically harmless and are not suitable for pharmaceutical applications but which can be used, for example, for the isolation or purification of the compounds of the invention.

Physiologically safe salts of the compounds of the invention include acid addition salts of mineral acids, carbonic acids and sulphonic acids, e.g. salts of hydrochloric acid, hydrobromic acid, sulphuric acid, phosphoric acid, methane sulphonic acid, ethanesulphonic acid, benzene sulphonic acid, toluene sulphonic acid, naphthalindisulphonic acid, acetic acid, trifluoroacetic acid, propionic acid, lactic acid, tartaric acid, malic acid, citric acid, fumaric acid, maleic acid and benzoic acid.

Physiologically safe salts of the compounds of the invention include salts of common bases, such as, and preferably, alkali metal salts (e.g. sodium and potassium salts), alkaline earth salts (e.g. calcium and magnesium salts) and ammonium salts derived from ammonia or organic amines with 1 to 16 C atoms, such as, and preferably, ethyllamine, diethylamine, triethylamine, ethyldiisopropylamine, monoethylamine, diethano, triethylamine, dicyclohexylamine, dimethylaminoethanol, procaine, dibenzylamine, n-methylpylamine, arginine, n-methylmethylphenylamine, 1,2-morphine, and l-ethylamine.

The solvates are defined as forms of the compounds of the invention which, in the solid or liquid state, form a complex by coordination with solvent molecules. Hydrates are a special form of solvates in which coordination with water takes place.

Err1:Expecting ',' delimiter: line 1 column 148 (char 147)

For the purposes of the present invention, the substitutes have the following meanings, unless otherwise specified: (C1-C6) alkyl and (C1-C4) alkyl represent a straight chain or branched alkyl residue with 1 to 6 and 1 to 4 carbon atoms respectively. A straight chain or branched alkyl residue with 1 to 4 carbon atoms is preferred. For example and preferably, the following are given: methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec.-butyl, tert-butyl, n-pentyl, 2-pentyl, 3-pentyl, neopentyl, n-hexyl, 2-hexyl and 3-hexyl.C1-C12) alkyl, (C1-6) alkylcandiol, (C1-C12) and (C2-C12) alkylcandiol (C6-C2) are given as a straight chain residue, with a mean of 12, 12 to 12, respectively, in the following example: α-alkyl, 1 to 6, 2 to 6 and 2 to 12 alkylcandiols.The use of a carbonyl cycloalkyl in the framework of the invention is called a monocyclic, interconnected carbonyl group with 3 to 6 carbon atoms. The two main examples of carbonyl cycloalkyl are cycloalkyl and cycloalkyl. The use of a carbonyl cycloalkyl is called a cycloalkyl, or cycloalkyl, is a pre-example of the invention. Cycloalkyl is a compound containing an alkyl group with 1 to 6 carbon atoms. Cycloalkyl is a chemical compound with two hydrocarbons. Cycloalkyl is a chemical compound with a hydroxy group and is a hydroxy group. Cycloalkyl is a chemical compound with a hydroxy group. Cycloalkyl is a chemical compound with a hydroxy group. Cycloalkyl is a chemical compound with a hydroxy group. Cycloalkyl is a chemical compound with a hydroxy group. Cycloalkyl is a chemical compound with a hydroxy group. Cycloalkyl is a chemical compound with a hydroxy group. Cycloalkyl is a chemical compound with a hydroxy group. Cycloalkyl is a chemical compound with a hydroxy group. Cycloalkyl is a chemical compound with a hydroxy group. Cycloalkyl is a chemical compound with a hydroxy group.The following are examples and preferences: methoxycarbonyl, ethoxycarbonyl, n-propoxycarbonyl, isopropoxycarbonyl, n-butoxycarbonyl and tert-butoxycarbonyl.Mono- ((C1-C4) alkylamine represents an amino group with a straight chain or branched alkyl substituent containing 1 to 4 carbon atoms. The following are examples and preferences: methylamine, ethylamine, n-propylamine, isopropylamine, n-butylamine and tert-butylamine.

For the purposes of the present invention, all residues occurring multiple times have a meaning independent of each other. If residues are substituted in the compounds of the invention, the residues may, unless otherwise specified, be substituted one or more times. A substitution with one or two identical or different substituents is preferred.

The preferred compounds for the present invention are those of formula (I) in which the R1 stands for hydrogen, (C1-C4) alkyl or a group of the formula Q1-L1-* or Q2-L2-* wherein Other * marks the point of attachment with the nitrogen atom, Q1 hydroxycarbonyl or (C1-C4) alkoxycarbonyl means: L1 straight chain (C1-C12) alkandiyl means that (a) two carbon atoms in a 1,3- or 1,4-relation can be broken into a phenyl ring by one or both of the carbon atoms between them, or (b) up to three non-adjacent CH2 groups can be replaced by -O, Q2 hydroxy, amino or methyllamine means and L2 straight chain (C2-C12) alkandiyl means that it can be substituted one or two times with methyl and that up to three non-adjacent CH2 groups can be replaced with -O-R2 stands for methyl or hydroxyR3 stands for hydrogenR4 stands for benzyl, 4-hydroxybenzyl, 1-phenylethyl or 1H-indol-3-ylmethyl or R3 and R4 together with the carbon atom to which they are both bound, form a 2-phenylcyclopropane-1,1-diyl group with the formula Other, including: # marks the junctions with the rest of the molecule, and the ring A with the N-O grouping contained therein for a monocyclic or bicyclic, where appropriate substituted heterocycle of the formula Other It says in **the linking point is marked with the carbonyl group, and R5 Hydrogen, hydroxy or benzyloxy means: and their salts, solvates and solvates of salts.

The present invention gives particular preference to compounds of formula (I) in which the R1 stands for hydrogen, methyl or a group with formula Q1-L1-* or Q2-L2-* where Other * marks the point of attachment with the nitrogen atom, Q1 Hydroxycarbonyl, methoxycarbonyl or ethoxycarbonyl means L1 straight chain (C1-C6) alkandiyl means that two carbon atoms can be broken into a phenyl ring in a 1,4-relation with the two carbon atoms between them, Q2 hydroxy or amino means and L2 straight chain (C2-C6) alkandiyl means,R2 is for methyl,R3 is for hydrogen,R4 is for benzyl, 1-phenylethyl or 1H-indol-3-ylmethyl, or R3 and R4 together with the carbon atom to which they are both bound, form a (1S,2R) -phenylcyclopropane-1,1-diyl group with the formula Other Other, including: #1the link with the adjacent nitrogen atom and #2the linking site with the carbonyl group, and the ring A with the N-O grouping contained therein for a monocyclic or bicyclic, where appropriate substituted heterocycle of the formula It says in **the linking point is marked with the carbonyl group, and R5 Hydrogen, hydroxy or benzyloxy means: and their salts, solvates and solvates of salts.

The compounds of formula (I-A) are of particular importance in the context of the present invention. Other where R1, R2, R3, R4 and ring A have the meanings defined above and the Cx carbon atom bearing the residues R3 and R4 has the S configuration shown, and their salts, solvates and solvates of salts.

The residue definitions given in the respective combinations or preferred residue combinations shall be replaced by residue definitions of other combinations as appropriate, irrespective of the respective combinations of residues given.

In particular, combinations of two or more of the above preferences are preferred.

The invention also relates to a process for the preparation of the compounds of formula (I) according to the invention, characterized by the use of a compound of formula (II) Other where R2 has the above meaning and PG stands for an amino group such as (9H-fluorescent-9-ylmethoxy) carbonyl, tert-butoxycarbonyl or benzyloxycarbonyl, in an inert solvent with activation of the carboxyl function in (II) either [A] first with the compound formula (III) Other or a salt thereof to a compound of formula (IV) Other where R2 and PG have the meanings given above, The product is then coupled and injected into an inert solvent by activation of the carboxyl function with a compound of formula (V) where R3, R4 and ring A have the meanings given above,Other or a salt of this compound to a compound of formula (VI) Other where R2, R3, R4, ring A and PG have the meanings given above, Other, of circular cross-section or[B] with a compound of formula (VII) Other where R3, R4 and ring A have the meanings given above, or a salt of this compound or of the compound with formula (VI) where R2, R3, R4, ring A and PG have the meanings given above, Other, of circular cross-section and the resulting compound of formula (VI) is then, by the usual methods of peptide chemistry, converted into a compound of formula (I-B) of the invention Other where R2, R3, R4 and ring A have the meanings given above,Other Protects and subsequently, if desired, either (i) by base-induced alkylation with a compound of formula (VIII) Other The following shall be used for the calculation of the maximum permissible mass: Other in which R1A has the above meaning of R1 but does not stand for hydrogen, and X stands for a volatile group such as chloride, bromide, iodide, mesylate, triphlate or tosylate, a width of not more than 50 mm, where R1A, R2, R3, R4 and ring A have the meanings given above, Translated or (ii) translated with a compound of formula (IX) Other in which R1B has the above value but in the alkyl chain length is reduced by one CH2 unit to R1A, in the presence of a suitable reducing agent in a compound of the formula (I-D) according to the invention Other in which R1B, R2, R3,R4 and ring A have the meanings given above, The Commission shall: and separates the resulting compounds of the formulae (I-B), (I-C) or (I-D) respectively, as appropriate, into their enantiomers and/or diastereomers and/or converts them into their solvates, salts and/or solvates of the salts with the appropriate (i) solvents and/or (ii) bases or acids.

The coupling reactions described above (amide formation from the respective amino and carbonic acid components) are carried out by commonly used methods of peptide chemistry (see, for example, M. Bodanszky, Principles of Peptide Synthesis, Springer-Verlag, Berlin, 1993; M. Bodanszky and A. Bodanszky, The Practice of Peptide Synthesis, Springer-Verlag, Berlin, 1984; H.-D. Jakubke and H. Jeschkeit, Amino acids, peptides, proteins, Verlag Chemie, Weinheim, 1982).

Inert solvents for the coupling reactions (II) + (III) → (IV), (IV) + (V) → (VI) and (II) + (VII) → (VI) are, for example, ethers such as diethyl ether, diisopropyl ether, tert-butyl methyl ether, tetrahydrofuran, 1,4-dioxane, 1,2-dimethoxyethane or bis- (((2-methoxyethyl) ether, hydrocarbons such as benzene, toluene, xylol, pentane, hexane, heptane, cyclohexane or petroleum fractions, halogenated hydrocarbons such as dichloromethane, trichloromethane, tetrahydromethane, 1,2-dichloromethane, trichloromethane or tetrahydrochloromethane, or mixed or dipolar solvents such as dimethyl nitrate, dimethyl nitrate, dimethyl nitrate, NMPN, dimethyl nitrate, dimethyl nitrate, NMPN (N), dimethyl nitrate, dimethyl nitrate, dimethyl nitrate, dimethyl nitrate, dimethyl nitrate, dimethyl nitrate, dimethyl nitrate, dimethyl nitrate, dimethyl nitrate, dimethyl nitrate, dimethyl nitrate, dimethyl nitrate, dimethyl nitrate, dimethyl nitrate, dimethyl nitrate, dimethyl nitrate, dimethyl nitrate, dimethyl nitrate, dimethyl nitrate, dimethyl nitrate, dimethyl nitrate, dimethyl nitrate, dimethyl nitrate, dimethyl nitrate, dimethyl nitrate, dimethyl nitrate, dimethyl nitrate, dimethyl nitrate, dimethyl nitrate, dimethyl nitrate, dimethyl nitrate, dimethyl nitrate, dimethyl nitrate, dimethyl nitrate, dimethyl nitrate, dimethyl nitrate, dimethyl nitrate, dimethyl nitrate, dimethyl nitrate, dimethyl nitrate, dimethyl nitrate, dimethyl nitrate, dimethyl nitrate,

For example, carbo-dihydrides such as N,N'-diethyl, N,N'-dipropyl, N,N'-diisopropyl, N,N'-dicyclohexylcarbodiamide (DCC) or N-dimethylaminopropyl) -N'-ethylcarbodiamide hydrochloride (EDC) are suitable as activating/condensing agents for these couplings; phlycyrogen derivatives such as N,N'-carbonyldiamidazole (CDI) or isobutyl chloride formate; 1,2-dioxyzolamide compounds such as 2-ethyl-5-phenyl-1,2-dioxyzolamine-3-sulphate or 2-tetrahydramethetrametyl-5-methyltetrahydrametrizol (THF-fluoro-1, N-dioxyzolamine), 2-dioxyzolamine, 2-dioxyzolamine, 2-dioxyzolamine, 2-dioxyzolamine, 2-dioxyzolamine, 2-dioxyzolamine, 2-dioxyzolamine, 2-dioxyzolamine, 2-dioxyzolamine, 2-dioxyzolamine, 2-dioxyzolamine, 2-dioxyzolamine, 1-dioxyzolamine, 2-dioxyzolamine, 1-dioxyzolamine, 1-dioxyzolamine, 1-dioxyzolamine, 1-dioxyzolamine, 1-dioxyzolamine, 1-dioxyzolamine, 1-dioxyzolamine, 1-dioxyzoyl or 2-dioxyzoyl-methyl or 2-dioxyzoyl-methyl-methyl-methyl, or 1-dioxyriphenyl-methyl-methyl-methyl, or 2-dioxylamine, or other compounds, such as N-dioxyzoyl-dioxy-dioxy-dioxy-dioxy-meth, 1-dioxy-meth, 2-dioxy-meth, 2-dioxy-methyl, 2-dioxy-methyl, 2-dioxy-methyl, 2-diox

For the purposes of the present invention, N- ((3-dimethylaminoisopropyl) N'-ethylcarbodiimide hydrochloride (EDC) is preferred as an activation/condensation agent for such coupling reactions in combination with 1-hydroxybenzotriazole (HOBt) and N,N-diisopropylethylamine, or O- ((7-azabenzotriazole-1-yl) N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU) also in combination with N,N-disopropylethylamine.

The coupling reactions (II) + (III) → (IV), (IV) + (V) → (VI) and (II) + (VII) → (VI) are usually carried out in a temperature range of -20°C to +60°C, preferably at 0°C to +40°C. The conversions can be carried out at normal, increased or reduced pressure (e.g. 0.5 to 5 bar); in general, normal pressure is used.

For example, ether such as diethyl ether, diisopropyl ether, methyl tert-butyl ether, tetrahydrofuran, 1,4-dioxane, 1,2-dimethoxyethane or bis-(2-methoxyethyl) ether, hydrocarbons such as benzene, toluene, xylol, pentane, hexane, heptane, cyclohexane or petroleum fractions, or dipolar non-protic solvents such as acetone, methyl ketone, acetonite, dimethylfoxide (DMSO), N,N-dimethylform (DMF), N,N-dimethylform (DMA), N'-dimethylform (NMP), N-dimethylpropylform (DMA), N-dimethylform (NMP), N-dimethylform (DMA), N-dimethylform (NMP), N-dimethylform (NMP), N-dimethylform (NMP), N-dimethylform (NMP), N-dimethylform (NMP), N-dimethylform (NMP), N-dimethylform (NMP), N-dimethylform (NMP) or N-dimethylform (NMP) may be used.

Suitable bases for this alkylation reaction are in particular alkali hydroxides such as lithium, sodium or potassium hydroxide, alkaline or alkaline earth carbonates such as lithium, sodium, potassium, calcium or caesium carbonate, or common organic amines such as triethylamine, N-methylmorpholine, N-methylpiperidine, N,N-diisopropylethylamine, pyridine or 4-N,N-dimethylamylamine. Potassium or caesium carbonate is preferred. If necessary, the addition of an allylation catalyst is advantageous, such as lithium bromide or -iodide, sodium or butadium bromide, potassium bromide or -iodide or -iodide or -iodide or -iodide.

The reaction (I-B) + (VIII) → (I-C) is generally carried out in a temperature range of -20°C to +60°C, preferably at 0°C to +40°C. The implementation can be carried out at normal, increased or reduced pressure (e.g. 0.5 to 5 bar); normally, normal pressure is used.

The implementation (I-B) + (IX) → (I-D) is carried out in the solvents inerted under the reaction conditions, which are common for reductive amination, where appropriate in the presence of an acid and/or a water-repellent agent as catalyst. Such solvents include, for example, alcohols such as methanol, ethanol, n-propanol, isopropanol, n-butanol or tert-butanol, ethers such as tetrahydrofuran, 1,4-dioxane, 1,2-dimethoxyethane or bis-dioxane, or other solvents such as dichloromethane, 1,2-dioxane, N,N-dimethylformamide or water. It is also possible to add this solvent. 1,4-dioxane is used as a catalyst before water or as a diluent of acidic acid.

Complex borohydrides, such as sodium borohydride, sodium cyanoborohydride, sodium triacethoxyborohydride or tetra-n-butylammonium borohydride, are particularly suitable as reducing agents for this reaction.

The reaction can be carried out at normal, increased or decreased pressure (e.g. 0.5 to 5 bar); normally, normal pressure is used.

In the above steps, if appropriate or necessary, functional groups, such as amino, hydroxy and carboxyl groups, may be present in the residues R1, R1A, R1B, R2, R4, R5 and/or R6 in temporary protected form, where appropriate, and such protective groups may be introduced and removed by conventional methods known from peptide chemistry (see, e.g., T.W. Greene and P.G.M. Wuts, Protective Groups in Organic Synthesis, Wiley, New York, 1999; M. Bodanszky and A. Bodanszky, The Practice of Peptide Synthesis, Springer-Verlag, Berlin, 1984).

The amino protective group is preferably tertiary butyric carbon (Boc), benzyl butyric carbon (Z) or (9H-fluorine-9-ylmethoxy) carbon (Fmoc); for a hydroxy or carboxyl function, tertiary butyl or benzyl is preferably used as protective group. The cleavage of a tertiary butyric carbon is usually done by treatment with a strong acid such as hydrochloric acid, bromohydrogen or trifluoric acid in an inert solvent such as diethyl fluoride, 1,4-D-methyl methoxy, dichloromethoric acid or benzoic acid; this reaction can also be carried out in an appropriate secondary addition to a solvent. In the case of benzyl butyric acid or benzyl butyric acid, such as piperyl or diethyl methoxy, this group is removed in the presence of a protective base such as diethyl methoxy or diethyl methoxy-9-methyl.

The compounds of formula (II) can be prepared by conventional peptide chemistry methods, for example by first taking N- ((benzyloxycarbonyl) -L-valine of formula (X) Other where Z stands for the benzyloxycarbonyl protective group, a thickness of not more than 0,05 mm, Other where T stands for (C1-C4) alkyl, or a salt of this compound to a compound of formula (XII) Other where T and Z have the meanings given above, The test chemical is then combined with the test chemical in the presence of a condensing agent containing N-protected N-methyl-L-valine or N-methyl-L-threonine of formula (XIII) where R2 has the above meaning and PG stands for an amino group such as (9H-fluorescent-9-ylmethoxy) carbonyl, tert-butoxycarbonyl or benzyloxycarbonyl, a purity by weight of more than 0,5% Other where R2, PG and T have the meanings given above, The test chemical is then combined with the ester group -C ((O) OT in (XIV) and transferred to the free carbonate by the usual methods.

The couplings (X) + (XI) → (XII) and Z-protected (XII) + (XIII) → (XIV) shall be performed under analogue reaction conditions as described in the coupling steps described in procedures [A] and [B].

The hydrolysis of the ester group -C(O)OT in step (XIV) → (II) is carried out by the usual method by treating the ester in an inert solvent with an acid or a base, in the latter case the resulting carboxylate salt is transferred to the free carbonate by the subsequent addition of an acid.

The alkyl residue T in the compound (XI) is selected in such a way that the conditions of its cleavage are compatible with the respective protective group PG from compound (XIII) used.

The usual inorganic bases are suitable as bases for ester hydrolysis, in particular alkaline or alkaline hydroxides such as lithium, sodium, potassium or barium hydroxide, or alkaline or alkaline carbonates such as sodium, potassium or calcium carbonate.

The ester cleavage acids are generally sulphuric acid, hydrochloric/ hydrochloric acid, hydrobromic/ hydrobromic acid, phosphoric acid, acetic acid, trifluoric acid, toluene sulphonic acid, methane sulphonic acid or trifluorimethanesulphonic acid or mixtures thereof, if appropriate, with the addition of water.

Inert solvents are water or organic solvents used for ester cleavage. These include preferably low alcohols such as methanol, ethanol, n-propanol or isopropanol, ethers such as diethyl ether, tetrahydrofuran, 1,4-dioxane or 1,2-dimethoxyethane, or other solvents such as dichloromethane, acetone, methyl ethyl ketone, N,N-dimethylformamide or dimethyl sulphide. Mixtures of these solvents may also be used. In the case of a basic ester hydrolysis, mixtures of water with 1,4-dimethylformamide, tetrahydrofuran, methanol, methanol and/or diethyl acid are preferred. In the case of the implementation, diethyl or 1,4-dimethyl sulphide is preferred.

The ester cleavage generally occurs in a temperature range of -20°C to +100°C, preferably 0°C to +50°C.

The compounds of formula (V) can be prepared by analogy with known processes, for example by taking a protected amino acid of formula (XV) Other where R3 and R4 have the meanings given above and Boc stands for tert-butoxycarbonyl protecting group, with activation of the carboxyl function either [C] with a compound of the formula (XVI) in which the ring A has the above meaning, or a salt of this compound to a compound of formula (XVII) Other where R3, R4, ring A and Boc have the meanings given above, Other, of circular cross-section or[D] first with hydroxylamine or a salt thereof to a compound of the formula (XVIII) Other where R3, R4 and Boc have the meanings given above, a thickness of not more than 0,05 mm, Other in which the connecting group A' corresponds to the other elements of the ring A as defined above, except the N-O grouping, if any, substituted, Cycling also with the compound formula (XVII) Other where R3, R4, ring A and Boc have the meanings given above, Alkylated and then the Boc protective group in (XVII) is broken down by the usual route by treatment with an acid.

The couplings (XV) + (XVI) → (XVII) and (XV) + hydroxylamine → (XVIII) shall be performed under analogue reaction conditions as described in the coupling steps described in procedures [A] and [B].

The base for cycloalkylation in step (XVIII) + (XIX) → (XVII) is preferably alkaline hydroxides such as lithium, sodium or potassium hydroxide or alkaline or alkaline earth carbonates such as lithium, sodium, potassium, calcium or caesium carbonate.

Inert solvents are particularly suitable for this reaction, such as alcohols such as methanol, ethanol, n-propanol, isopropanol, n-butanol or tert-butanol, or dipolar-aprotic solvents such as acetone, methylethyl ketone, N,N-dimethylformamide or dimethyl sulfoxide.

The reaction can be carried out at normal, increased or decreased pressure (e.g. 0.5 to 5 bar); normally, normal pressure is used.

The compounds of formula (VII) are in turn obtained by coupling the above compound (V) to the compound (XX) in which box is the tert-butoxycarbonyl protection group, to a compound of formula (XXI) Other where R3, R4, ring A and Boc have the meanings given above, The compound (XX) itself is available from compound (III) via a corresponding protective group introduction.

The coupling reaction (V) + (XX) → (XXI) shall again be carried out under the same conditions as described in the coupling steps described in methods [A] and [B].

The compounds of formulae (III), (VIII), (IX), (X), (XI), (XIII), (XV), (XVI) and (XIX), including, where appropriate, chiral or diastereomeric forms thereof, are commercially available or described as such in the literature, or may be prepared by means obvious to the professional by analogy with methods published in the literature.

If appropriate isomer-pure starting materials are not available, separation of the compounds of the invention into the corresponding enantiomers and/or diastereomers may be conveniently carried out at the stage of the compounds (V), (VI), (VII), (XV), (XVI), (XVII), (XVIII) or (XXI), which may then be further separated in the reaction steps described. Such separation of the stereoisomers may be carried out by the usual methods known to the skilled. Chromatographic methods may be preferred on achiric or chiral bases; in the case of free carbon acids, as an alternative intermediate, separation by chiral diastereomers may also be carried out by means of saline bases.

The manufacture of the compounds of the invention can be illustrated by the following reaction schemes: Other

The compounds of the invention have valuable pharmacological properties and can be used for the prevention and treatment of diseases in humans and animals.

The compounds of the invention have significantly higher cytotoxic activity compared to monomethylauristatin F (MMAF) and significantly increased plasma stability compared to known ester derivatives of MMAF such as MMAF-OMe. The compounds of the invention are therefore particularly suitable for the treatment of hyperproliferative diseases in humans and mammals in general because of this property profile. The compounds can inhibit, block, decrease or reduce cell proliferation and cell division and enhance apoptosis.

The hyperproliferative diseases for which the compounds of the invention may be used include, in particular, the group of cancer and tumour diseases, including, but not limited to, the following diseases for the purposes of the present invention: breast cancer and breast tumours (ductal and lobular forms, including in situ), respiratory tumours (small and non-small cell carcinoma, bronchial cancer), brain tumours (e.g. brainstem and hypothalamus, astrocytoma, medulloblastoma, ectodermal and neuroectodermal tumours), tumours of the organs of the body (spleen, spleen, lipoma, gallbladder, thyroid, spleen, spleen, spleen, spleen, spleen, spleen, spleen, spleen, spleen, spleen, spleen, spleen, spleen, spleen, spleen, spleen, spleen, spleen, spleen, spleen, spleen, spleen, spleen, spleen, spleen, spleen, spleen, spleen, spleen, spleen, spleen, spleen, spleen, spleen, spleen, spleen, spleen, spleen, spleen, spleen, spleen, spleen, spleen, spleen, spleen, spleen, spleen, spleen, spleen, spleen, spleen, spleen, spleen, spleen, spleen, spleen, spleen, spleen, spleen, spleen, spleen, spleen, spleen, spleen, spleen, spleen, spleen, spleen, spleen, spleen, spleen, spleen, spleen, spleen, spleen, spleen, spleen, spleen, spleen, spleen, spleen, spleen, spleen, spleen, spleen, spleen, spleen, spleen, spleen, spleen, spleen, spleen, spleen, spleen, spleen, spleen, spleen, spleen, spleen, spleen, spleen, spleen, spleen, spleen, spleen, spleen, spleen, spleen, spleen, spleen, spleen, spleen, spleen, spleen, spleen, spleen, spleenThe risk of infection is high in patients with Kaposi' s sarcoma, malignant melanoma, Merkel cell and non-melanoma skin cancers, tumours of the soft tissue (including soft tissue, osteosarcoma, malignant fibrous histiocytoma, lymphomas and rhabdomyosarcoma), tumours of the eyes (including intraocular melanoma and retinoblastoma), tumours of the endocrine and exocrine glands (e. g. thyroid and parathyroid glands, pancreatic and salivary glands), tumours of the urinary tract (including bladder, penile, kidney, renal and urinary cysts), and of the reproductive organs (including endometriosis, vaginal, vulvar, and ovarian cancer, acute myelogenous and prostatic cancer, and chronic myelogenous leukemia), including acute forms of cancer and prostate cancer, and in patients with acute myelogenous and prostate cancer, including prostate cancer, prostate cancer, prostate cancer and prostate cancer, and chronic myelogenous leukemia, and prostate cancer, including cancer of the prostate, prostate and prostate glands, and prostate glands, and prostate glands, and prostate glands, and prostate glands, and prostate glands, and prostate glands).The following conditions are considered to be relevant for the diagnosis of Hodgkin lymphoma:

These well-described human diseases can also occur in other mammals with similar etiology and be treated there with the compounds of the present invention.

Err1:Expecting ',' delimiter: line 1 column 55 (char 54)

The present invention is thus also concerned with the use of the compounds of the invention for the treatment and/or prevention of diseases, in particular the diseases mentioned above.

The present invention also relates to the use of the compounds of the invention for the manufacture of a medicinal product for the treatment and/or prevention of diseases, in particular the diseases mentioned above.

The present invention also relates to the use of the compounds in a process for the treatment and/or prevention of diseases, in particular the diseases mentioned above.

The present invention also relates to a method for the treatment and/or prevention of diseases, particularly those mentioned above, using an effective amount of at least one of the compounds of the invention.

The compounds of the invention may be used alone or, if necessary, in combination with one or more other pharmacologically active substances, provided that such combination does not lead to undesirable and unacceptable side effects.

For example, the compounds of the present invention may be combined with known antihyperproliferative, cytostatic or cytotoxic substances for the treatment of cancer, for example:

The following are the active substances which are to be classified in the additive: - aldoxyurea, aldoxyurea, aldoxyurea, aldoxyurea, aldoxyurea, aldoxyurea, aldoxyurea, aldoxyurea, aldoxyurea, aldoxyurea, aldoxyurea, aldoxyurea, aldoxyurea, aldoxyurea, aldoxyurea, aldoxyurea, aldoxyurea, aldoxyurea, aldoxyurea, aldoxyurea, aldoxyurea, aldoxyurea, aldoxyurea, aldoxyurea, aldoxyurea, aldoxyurea, aldoxyurea, aldoxyurea, aldoxyurea, aldoxyurea, aldoxyurea, aldoxyurea, aldoxyurea, aldoxyurea, aldoxyurea, aldoxyurea, aldoxyurea, aldoxyurea, aldoxyurea, aldoxyurea, aldoxyurea, aldoxyurea, aldoxyurea, aldoxyurea, aldoxyurea, aldoxyurea, aldoxyurea, aldoxyurea, aldoxyurea, aldoxyurea, aldoxyurea, aldoxyurea, aldoxyurea, aldoxyurea, aldoxyurea, aldoxyurea, aldoxyurea, aldoxyurea, aldoxyurea, aldoxyurea, aldoxyurea, aldoxyurea, aldoxyurea, aldoxy, aldoxy, aldoxy, aldoxy, aldoxy, aldoxy, aldoxy, aldoxy, aldoxy, aldoxy, aldoxy, aldoxy, aldoxy, aldoxy, aldoxy, aldoxy, aldoxy, aldoxy, aldoxy, aldoxy, aldoxy, aldoxy, aldoxy, aldoxy, aldoxy, aldoxy, aldoxy, aldoxy, aldoxy, aldoxy, aldoxy, aldoxy, aldoxy, aldoxy, aldoxy, aldoxy, aldoxy, aldoxy, aldoxy, aldoxy, aldoxy, aldoxy, aldoxy, aldoxy, alThe following substances, which are to be classified in this class, are: gliadel, goserelin, granisetron hydrochloride, histrelin, hycamtin, hydrocorton, erythro-hydroxynonyladenine, hydroxyurea, ibritumomab tiuxetan, idarubicin, ifosfamide, interferon alpha, interferon alpha-2, interferon alpha-2a, interferon alpha-2β, interferon alpha-n1, interferon alpha-n3, interferon beta, interferon gamma-1a, interleukin-2, intron A, irressa, irinotecan, kytramil, lentomanan sulphate, letrozone, leucovorin, leuproxydrin, leuproximide acetate, premyloxazine, leuproxyde citrate, levoplatin, leucotrizin, leucotrizin, leucotrizin, leucotrizin, leucotrizin, leucotrizin, leucotrizin, leucotrizin, leucotrizin, leucotrizin, leucotrizin, leucotrizin, leucotrizin, leucotrizin, leucotrizin, leucotrizin, leucotrizin, leucotrizin, leucotrizin, leucotrizin, leucotrizin, leucotrizin, leucotrizin, leucotrizin, leucot, leucotrizin, leucotrin, leucotrin, leucotrin, leucotrin, leucotrin, leucrozin, leucrozin, leucrozin, leucrozin, leucrozin, leucrozin, leucro, leucro, leucro, leucro, leucro, leucro, leucro, leucro, leucro, leucro, leucro, leucro, leucro, leucro, leucro, leucro, leucro, leucro, leucro, leucro, leucro, leucro, leucro, leucro, leucro, leucro, leucro, leThe additive and premixtures, whether or not containing added sugar or other sweetening matter, are to be classified in the same category as the additive and premixtures, provided that the total value of the additive and premixtures is equal to or greater than the corresponding amount of the active substance, and the total value of the active substance, calculated on the basis of the additive and premixtures, is not more than the corresponding amount of the active substance.The active substance is a mixture of the following compounds: 13-cis-retinoic acid, satraplatin, seocalcitol, T-138067, tarceva, taxoprexin, thymosin alpha-1, thiazofurin, tipifarnib, tirapazamine, TLK-286, toremifene, TransMID-107R, valspodar, vapreotide, vatalanib, verteporfin, vinflunin, Z-100, zoledronic acid, and any combination of these.

In a preferred embodiment, the compounds of the present invention may be combined with antihyperproliferative agents, which may be, without being exhaustive, by way of example:

The following substances, which are not classified in this class, are to be classified in the same subcategory as the active substance: - glucosamine, - L-asparaginase, - azathioprine, - 5-azacytidine, - bleomycin, - busulfan, - carboplatin, - carmustin, - chlorambucil, - cisplatin, - colaspase, - cyclophosphamide, - cytarabin, - dacarbazine, - dactinomycin, - daunorubicin, - diethylstilbestrol, - 2',2'-difluorooxycytidine, - docetaxel, - doxorubicin (adriprocin), - epirubicin, - ehilon and its derivatives, - erythro-hydroxynonyladenine, - ethylstradiol, - tamoxifen, - chlorambucil, - collagen, - collagen, - collagen, - collagen, - collagen, - collagen, - collagen, - collagen, - collagen, - collagen, - collagen, - collagen, - collagen, - collagen, - collagen, - collagen, - collagen, - collagen, - collagen, - collagen, - collagen, - collagen, - collagen, - collagen, - collagen, - collagen, - collagen, - collagen, - collagen, - collagen, - collagen, - collagen, - collagen, - collagen, - collagen, - collagen, - collagen, - collagen, - collagen, - collagen, - collagen, - collagen, - collagen, - collagen, - collagen, - collagen, - collagen, - collagen, - collagen, - collagen, - collagen, - collagen, - collagen, - collagen, - collagen, - collagen, - collagen, - collagen, - collagen, - collagen, - collagen, - collagen, - collagen, - collagen, - collagen, - collagen, - collagen, - collagen, - collagen, - collagen, - collagen, - collagen, - collagen, - collagen, - collagen

The compounds of the invention can also be promisingly combined with biological therapeutics such as antibodies (e.g. Avastin, Rituxan, Erbitux, Herceptin). The compounds of the invention can also be used in combination with antiangiogenesis therapies such as Avastin, Ainibxit, Recentin, Regorafenib, Sorafenib or Sunitinib to achieve positive effects. Combinations with proteasome inhibitors and mTOR inhibitors, as well as combinations with antihormones and steroidal metabolic enzyme inhibitors are also particularly suitable due to their favourable side effects profile.

In general, the following objectives can be achieved by combining compounds of the present invention with other agents having cytostatic or cytotoxic activity: The results of the study showed that the use of chemotherapy in the treatment of cancer was more effective in slowing tumour growth, reducing tumour size or even eliminating tumour growth compared to treatment with a single active substance;the possibility of using the chemotherapeutic agents used at a lower dose than monotherapy;the possibility of using a more tolerable therapy with fewer side effects compared to single administration;the possibility of treating a wider range of tumour diseases;the achievement of a higher response rate to the therapy;a longer survival of patients compared to today's standard therapy.

In addition, the compounds of the invention may be used in combination with radiation therapy and/or surgery.

The present invention also covers medicinal products containing at least one compound of the invention, usually together with one or more inert non-toxic excipients suitable for pharmaceutical use, and their use for the purposes described above.

The compounds of the invention may act systemically and/or locally and may be applied in appropriate ways, such as orally, parenterally, pulmonally, nasally, sublingually, lingually, buccally, rectally, dermally, transdermally, conjunctivally, ottically or as an implant or stent.

For these applications, the compounds of the invention may be administered in suitable application forms.

For oral application, the fast and/or modified forms of application that contain the compounds of the invention in crystalline and/or amorphous and/or dissolved forms, such as tablets (uncoated or coated tablets, for example with stomach-resistant or delayed dissolving or insoluble preparations that control the release of the compounds of the invention), or fast-decay tablets or films/oils, films/lyophilisates, capsules (for example, viscose or soft capsules), granules, emulsions, pulses, solutions, aerosols, or suspensions, in the oral cavity, are suitable according to the state of the art.

Parenteral administration may be bypass an absorption step (e. g. intravenous, intra-arterial, intra-cardial, intraspinal or intralumbar) or include absorption (e. g. intramuscular, subcutaneous, intracutan, percutan or intraperitoneal).

Other routes of application include inhalation formulations (including powder inhalers, nebulizers), nasal drops, solutions or sprays, tablets, films/foils or capsules for oral, sublingual or oral application, suppositories, ear or eye preparations, vaginal capsules, aqueous suspensions (lotions, shakes), lipophilic suspensions, ointments, creams, transdermal therapeutic systems (e.g. plums), milk, pastes, foams, spray powders, implants or stents.

Oral or parenteral administration, particularly oral and intravenous administration, is preferred.

The compounds of the invention may be converted into the above applications by means of known in-house mixtures with inert, non-toxic, pharmaceutically suitable excipients, including carriers (e.g. microcrystalline cellulose, lactose, mannitol), solvents (e.g. liquid polyethylene glycols), emulsifiers and dispersing or net agents (e.g. sodium dodecyl sulphate, polyoxysorbitanoleate), binders (e.g. polyvinylpyrrolidone), synthetic and natural polymers (e.g. albumin), stabilizers (e.g. antioxidants), ascorbic acid (e.g. ascortic acid) and synthetic pigments (e.g. albioxide and albioxide).

In general, it has been shown to be beneficial to administer parenteral doses of approximately 0.001 to 1 mg/kg, preferably 0.01 to 0.5 mg/kg body weight, for effective results; oral doses of approximately 0.01 to 100 mg/kg, preferably 0.01 to 20 mg/kg, and particularly preferably 0.1 to 10 mg/kg body weight.

However, it may be necessary to deviate from the amounts mentioned, depending on body weight, route of application, individual behaviour with the active substance, type of preparation and time or interval of application. In some cases, it may be sufficient to use less than the minimum amount mentioned above, while in other cases the upper limit mentioned above may have to be exceeded.

The following examples of execution illustrate the invention. The invention is not limited to the examples.

The percentages in the following tests and examples are percentages by weight, parts by weight, solvent ratios, dilution ratios and liquid/liquid solution concentration ratios are all related to volume.

A. Examples Abbreviations and acronyms:

The following is a list of the active substances which may be used in the preparation of the active substance: acetylated acetylated acetylated acetylated acetylated acetylated acetylated acetylated acetylated acetylated acetylated acetylated acetylated acetylated acetylated acetylated acetylated acetylated acetylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acylated acy

HPLC and LC-MS methods: The test chemical is used to determine the concentration of the test substance.

The test method is based on the following equation:

The test chemical is used to determine the concentration of the test substance.

The test method is based on the following equation:

The test chemical is used to determine the concentration of the test substance.

The instrument is equipped with a micromass Quattro Micro MS with HPLC Agilent series 1100; column: Thermo Hypersil GOLD 3μ 20 mm x 4 mm; Eluent A: 1 1 water + 0.5 ml 50% formic acid, Eluent B: 1 1 acetonitrile + 0.5 ml 50% formic acid; gradient: 0.0 min 100% A → 3.0 min 10% A → 4.0 min 10% A → 4.01 min 100% A (flow 2.5 ml/min) → 5.00 min 100% A; oven: 50°C; flow: 2 ml/min; UV detection: 210 nm.

The test chemical is used to determine the concentration of the test substance.

The test chemical is used to determine the concentration of the active substance in the test chemical and to determine the concentration of the active substance in the test chemical.

The test chemical is used to determine the concentration of the test substance.

The device is HP 1090 series II; the column is Merck Chromolith SpeedROD RP-18e, 50 mm x 4.6 mm; the front column is Merck Chromolith Guard Cartridge Kit RP-18e, 5 mm x 4.6 mm; the volume of the injection is 5 μl; the eluent A is 70% HClO4 in water (4 ml/ l), the device is B: acetonitrile; the gradient is 0.00 min 20% B → 0.50 min 20% B → 3.00 min 90% B → 3.50 min 90% B → 3.51 min 20% B → 4.00 min 20% B; the flow is 5 ml/ min; the column temperature is 40°C.

The test chemical is used to determine the concentration of the test substance.

The device is equipped with a water-based anti-fouling system, which is equipped with a water-based anti-fouling system.

The test chemical is used to determine the concentration of the test substance.

The test chemical is used to determine the concentration of the active substance in the test chemical and to determine the concentration of the active substance in the test chemical.

The test chemical is used to determine the concentration of the test substance.

The test chemical is used to determine the concentration of the active substance in the test chemical and to determine the concentration of the active substance in the test chemical.

The test chemical is used to determine the concentration of the test substance.

The test chemical is used to determine the concentration of the active substance in the test chemical and to determine the concentration of the active substance in the test chemical.

All reagents or reagents not explicitly described below are considered to have been commercially sourced from publicly available sources; for all other reactants or reagents not described below that were not commercially available or were sourced from sources not explicitly described below, a reference to the published literature describing their production is provided.

Primary compounds and intermediates: The output connection 1 It consists predominantly of hydrocarbons having carbon numbers predominantly in the range of C1 through C4.]

The title compound can be produced in various ways according to literature requirements, e.g. see Pettit et al., Synthesis 1996, 719; Shioiri et al., Tetrahedron Lett. 1991, 32, 931; Shioiri et al., Tetrahedron 1993, 49, 1913; Koga et al., Tetrahedron Lett. 1991, 32, 2395; Vidal et al., Tetrahedron 2004, 60, 9715; Poncet et al., Tetrahedron 50, 1994, 5345 It was produced here either as a free acid (as shown) or as the corresponding dicyclohexylamine salt.

Output connection 2 It consists predominantly of hydrocarbons having carbon numbers predominantly in the range of C1 through C5.]

The title link can be established in various ways according to literature, see for example Pettit et al., J. Org. Chem. 1994, 59, 1796; Koga et al., Tetrahedron Lett. 1991, 32, 2395; Shioiri et al., Tetrahedron Lett. 1991, 32, 931; Shioiri et al., Tetrahedron 1993, 49, 1913.

The output connection 3 N-α- (tert-butyric-carbonyl) -N-hydroxy-L-phenylalanamide

The title link was prepared in accordance with literature requirements (A. Ritter et al., J. Org. Chem. 1994, 59, 4602). The following information is provided for the purpose of the calculation of the yield: The test chemical is used to determine the concentration of the test substance in the test medium.

The output connection 4 1,2-Oxazolidine hydrochloride

The title link can be made according to literature requirements, see e.g. H. King, J. Chem. Soc. 1942, 432; it is also commercially available.

The output connection 5 1,2-Oxazinane hydrochloride

The title link can be made according to literature, see e.g. H. King, J. Chem. Soc. 1942, 432.

The output connection 6 The following substances are to be classified in the same category as the active substance:

The title compound can be produced in Boc-protected form according to literature prescriptions (see e.g. C. Johnson et al., Tetrahedron Lett. 1998, 39, 2059); the usual method of excretion was treatment with Trifluoric Acid and subsequent neutralization.

The yield is 149 mg (89% by weight).

The output connection 7 1,2-Oxazolidine-4-ol

The title link can be made according to literature standards, see for example N. Amlaiky, Synthesis 1982, 5, 426.

The output connection 8 It consists predominantly of hydrocarbons having carbon numbers predominantly in the range of C1 through C5.]

The title compound was prepared in accordance with a literature prescription (A. Ritter et al., J. Org. Chem. 1994, 59, 4602) from commercially available (1S,2R)-1-[tert-butoxycarbonyl) amino]-2-phenylcyclopropanocarboxylic acid (C. Cativiela et al., Chirality 1999, 11, 583).

The yield is 339 mg (59% by weight)

The test chemical is used to determine the concentration of the test substance in the test medium.

Intermediate 1 The following shall be indicated in the column for the product:

HP HP were obtained by dissolving 425 mg (1.7 mmol) N-[benzyloxy) carbonyl]-L-valine in 50 ml DMF and then 500 mg (1.7 mmol) tert-butyl- ((3R,4S,5S) 3-methoxy-5-methyl-4-methylamino) heptanoate hydrochloride (parent compound 2), 356 mg (1.9 mmol) 1-(3-dimethylaminopropyl) 3-ethylcarbodiesodium hydrochloride, 285 mg (1.9 mmol) 1-hydroxy-1H-benzotriazole hydrate and 655 mg (5.1 mmol) N-dihydroxy-diisopropyl nitrogenoate. The mixture was dissolved at 20 h. Subsequently, 90 mg (0.5 mmol) 1-dimethylaminopropyl nitrate (0.9 mmol) was dissolved in 120 mg (1-dimethylaminoethyl nitrogenoyl nitrate) and then treated with a mixture of 119 mg (1-dimethylaminoethyl nitrate) and 120 mg (1-dimethylaminoethylaminoethyl nitrate) (1-methylaminoethyl nitrate) (1-dihydroxylamino-methyl nitrate) and 120 mg (1-dihydroxylamino-diethyl nitrate) (1-diethyl nitrate) and then treated with a solution of 120 mg (1-dimethyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl) and 120 mg (1-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl) and a solution of 120 mg (1-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl) and a solution of 120 mg (1-methyl-methyl-methyl-methyl-methyl

The test chemical is then applied to the test chemical.

The test chemical is used to determine the concentration of the test substance in the test medium.

Intermediate 2 It consists predominantly of hydrocarbons having carbon numbers predominantly in the range of C1 through C5.]

500 mg (1 mmol) tert-butyl- ((3R,4S,5S) -4-[{N-[benzyloxy) carbonyl]-L-valyl} ((methyl) amino]-3-methoxy-5-methylheptanoate (intermediate 1) were dissolved in 50 ml of methanol and after adding 100 mg 10% palladium to activated carbon were hydrated for 1 h at RT at normal pressure. The catalyst was then filtered and the solvent removed in a vacuum. 370 mg (quant.) of the title compound was obtained as a nearly colourless oil.

The test chemical is then applied to the test chemical.

The test chemical is used to determine the concentration of the test substance in the test medium.

Intermediate 3 The substance is to be classified in the immediate vicinity of the product.

396 mg (1.1 mmol) N-[9H-fluorescent-9-ylmethoxy) carbonyl]N-methyl-L-valine were dissolved in 20 ml DMF and mixed successively with 365 mg (1 mmol) tert-butyl- ((3R,4S,5S) -methoxy-5-methyl-4-[methyl(L-valyl) amino]heptanoate (intermediate 2), 234 mg (1.2 mmol) 1-(3-dimethylaminopropyl) 3-ethylcarbodiimidide hydrochloride and 187 mg (1.2 mmol) 1-hydroxy-1H-benzotriazole hydrate. The mixture was stirred overnight at RT. The mixture was then further reduced to a mixture of semi-organic chlor-ammonic acid-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L-L

The yield is 660 mg (68% d.j.)

The test chemical is then applied to the test chemical.

The test chemical is used to determine the concentration of the test substance in the test medium.

Intermediate 4 The following substances are to be classified in the same heading as the active substance:

650 mg (0.94 mmol) N-[(9H-fluorescent-9-ylmethoxy) carbonyl]-N-methyl-L-valyl-N-[(3R,4S,5S)-1-tert-butoxy-3-methoxy-5-methyl-1-oxoheptan-4-yl]-N-methyl-L-valinamide (intermediate 3) were taken in 5 ml of dichloromethane, added to 5 ml of trifluoroacetic acid and stirred overnight in RT.

The test chemical is then applied to the test chemical.

The test chemical is used to determine the concentration of the test substance in the test medium.

Intermediate 5 It consists predominantly of hydrocarbons having carbon numbers predominantly in the range of C1 through C5.]

500 mg (1.9 mmol) N-ter-butoxycarbonyl) L-phenylalanine was dissolved in 10 ml DMF and stirred successively with 466 mg (3.8 mmol) 1,2-oxyzanine hydrochloride (parent compound 5), 433 mg (2.3 mmol) 1-3-dimethylaminopropyl) 3-ethylcarbodiimide hydrochloride, 382 mg (2.8 mmol) 1-hydroxy-1H-benzotriazole hydrate and 731 mg (5.7 mmol) N,N-diisopropyl. The mixture was stirred overnight in RT. The solution was then compressed into a mixture of semi-saturated ammonium chloride solution and ethylene glycol. The organic phase was separated, dissolved with sodium and magnesium sulphate solution (98%), and dissolved in a 620 mg solution of sodium and magnesium sulphate.

The test chemical is then applied to the test chemical.

The test chemical is used to determine the concentration of the test substance in the test medium.

Intermediate 6 It consists predominantly of hydrocarbons having carbon numbers predominantly in the range of C1 through C5.]

620 mg (1.85 mmol) tert-butyl[(2S)-1-(1,2-oxazinan-2-yl)-1-oxo-3-phenylpropan-2-yl]carbamat (intermediate 5) were added to 5 ml of dichloromethane, added to 10 ml of trifluoroacetic acid and stirred for 30 min at RT. The residue was then vacuum-pressed and lyophilised from water/acetonitrile to obtain 750 mg of the title compound as a colourless foam.

The test chemical is then applied to the test chemical.

The test chemical is used to determine the concentration of the test substance in the test medium.

Intermediate 7 It consists predominantly of hydrocarbons having carbon numbers predominantly in the range of C1 through C5.]

The organic phase was dried, filtered and solidified with magnesium sulphate. The residue was collected in 10 ml DMF and subsequently purified with 49 mg (0.11 mmol) (2S) 2-methylenedioxyran-2-yl) 3-phenyl-1-methoxypropanone tetrafluoroacetic acid (Intermediate), 61 mg (0.16 mmol) (61-methylenedioxyran-1-yl) tetrafluoroacetate (Intermediate), 61 mg (0.16 mmol) (64-methylenedioxyran-1-yl) tetrafluoroacetate (Intermediate), 56 mg (32-methylenedioxyran-1-yl) tetrafluoroacetate (Intermediate), and then with the intermediate tetrafluoroacetate (Intermediate), 61 mg (0.16 mmol) (61-methylenedioxyran-1-yl) tetrafluoroacetate (Intermediate), 56 mg (32-methylenedioxyran-1-yl) tetrafluoroacetate (Intermediate), 61 mg (0.16 mmol) (63-methylenedioxyran-1-yl) tetrafluoroacetate (Nitrate) was then re-satured with the intermediate tetrafluoroacetate tetrafluoroacetamethyl-1-yl-methyranolone (N-methylenedioxylenedioxyran-1-yl) tetrafluoroacetate (N-methylenedioxyran-2-yl) tetrafluoroacetyl) and 56 mg (32-methylenedioxylenedioxyran-1-yl) tetrafluoroacetamethyl) tetrafluoroacetate (N-methylenedioxyranolone) (1-methyranolone) (1-methylenedioxyranolone) (1-methyranolone) (1-methylenedioxylenedioxyran-2-methyran-2-yl) was then reduced to 56 mg (2-methylenedioxylenedioxyran-1-methyran-1-methyran-1-methyran-1-yl) tetrafluoroacet (N-methylenedioxyranolone) (1-methyranolone) (1-methy

The test chemical is then applied to the test chemical.

The test chemical is used to determine the concentration of the test substance in the test medium.

44 mg (0.09 mmol) of this intermediate was absorbed into 3 ml of dichloromethane, added to 1 ml of trifluoroacetic acid and stirred for 30 min at RT. The remaining residue was then vacuum-pressed and lyophilized from water/acetonitrile, resulting in 39 mg (86% dT) of the title compound as a colourless foam.

The test chemical is then applied to the test chemical.

The test chemical is used to determine the concentration of the test substance in the test medium.

Intermediate 8 It consists predominantly of hydrocarbons having carbon numbers predominantly in the range of C1 through C4.]

The mixture was stirred for 1 hour. The solvent was then removed in a vacuum, the residue was absorbed in ethyl acetate and then shaken with 5% citric acid solution and then with 5% sodium tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary tertiary terti

The test chemical is used to determine the concentration of the test substance in the test medium.

64 mg (0.18 mmol) of this intermediate was absorbed into 10 ml of dichloromethane, added to 1 ml of trifluoroacetic acid and stirred for 30 min at RT. The remaining water/dioxane residue was then vacuum-pressed and lyophilised to yield 66 mg (quant.) of the title compound as a colourless foam.

The test chemical is then applied to the test chemical.

The test chemical is used to determine the concentration of the test substance in the test medium.

Intermediate 9 It consists predominantly of hydrocarbons having carbon numbers predominantly in the range of C1 through C5 and boiling in the range of approximately -15oC to -15oC (-70oF to -15oF).]

The organic phase was first dried, filtered and solidified with magnesium sulphate, the residue was incorporated into 10 ml DMF and subsequently treated with 66 mg (0.18 mmol) (2S) 2-methylenedioxy-2-methylenedioxy-3-azabicarbonate (0.2.2) 3-methylenedioxy-3-phenylpropanedioxy-3-fluorooxaxa-acetate (0.2.2) 3-methylenedioxy-3-fluorooxa-acetate (0.2.2) 3-methylenedioxy-3-phenylpropanedioxy-3-fluorooxa-acetate (0.2.2) 3-methylenedioxylenedioxy-3-acetate (0.2.2) 8 mmol (N2O2-N2O2-methylenedioxy-3-acetate) (52 mmol) was then re-substituted with 52 mg (52.6 mmol) of mannitrile-1-methylenedioxy-1-methylenedioxy-1-acetate (52.2) 3-methylenedioxylenedioxy-3-acetylpropanedioxy-3-acetylpropanedioxy-3-acetate (0.2.2) 3-methylenedioxylenedioxy-3-phenylpropanedioxy-3-acet (32.2) 3-methylenedioxylenedioxy-3-acet (32.2) 3-methylenedioxylenedioxylenedioxy-3-acet (32.2) (N2O2-N2O2-N2O2-N2O2-N2-N2O3-acetylpropanedioxylenedioxy-3-acet) (52.2) 3-methylenedioxylenedioxy-3-acet) was then re-substituted with 52 mg (52.2) 53 mg (52.2) 2-methylenedioxylenedioxylenedioxylenedioxy-acet (32-methylenedioxy-acet (32-acet) and 2-methylenedioxylenedioxylenedioxy-acet (32-methylenedioxy-acet) 2-methylenedioxy-acet (32-methylenedioxy-

The test chemical is then applied to the test chemical.

The test chemical is used to determine the concentration of the test substance in the test medium.

52 mg (0.1 mmol) of this intermediate was absorbed into 10 ml of dichloromethane, added to 1 ml of trifluoric acid and stirred for 20 min at RT. Then the residual was vacuum-pressed and stirred with 20 ml of diethyl ether. After 10 min the filter was filtered and the filter residual was dried in a high vacuum, thus producing 39 mg (72% dT) of the title compound.

The test chemical is then applied to the test chemical.

The test chemical is used to determine the concentration of the test substance in the test medium.

Intermediate 10 It consists predominantly of hydrocarbons having carbon numbers predominantly in the range of C1 through C5.]

Initially, the Boc-protected intermediate tert-butyl-[(((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((

The following information is provided for the purpose of the calculation of the yield:

The test chemical is used to determine the concentration of the test substance in the test medium.

36 mg (0.094 mmol) of this intermediate was absorbed into 5 ml of dichloromethane, added to 0.5 ml of trifluoric acid and stirred for 20 min at RT. The residues were then vacuum-pressed and purified by HPLC. The corresponding fractions were combined, concentrated in a vacuum to a volume of about 15 ml and then lyophilised. 16 mg (43% dT) of the titanium compound was obtained as a colourless foam.

The test chemical is then applied to the test chemical.

The test chemical is used to determine the concentration of the test substance in the test medium.

Intermediate 11 It consists predominantly of hydrocarbons having carbon numbers predominantly in the range of C1 through C5 and boiling in the range of approximately -15oC to -15oC (-70oF to -15oF).]

The organic phase was first dried, filtered and ejected via magmus sulphate. The residue was collected in 10 ml DMF and subsequently treated with 16 mg (0.031 mmol) (2S) 2-methylenedioxy-1-methylenedioxy-3-methylenedioxy-3-methyl-3-methyl-3-methyl-3-methyl-3-methyl-3-methyl-3-methyl-3-methyl-3-methyl-3-methyl-3-methyl-3-methyl-3-methyl-3-methyl-3-methyl-3-methyl-3-methyl-3-methyl-3-methyl-3-methyl-3-methyl-3-methyl-3-methyl-3-methyl-3-methyl-3-methyl-3-methyl-3-methyl-3-methyl-3-methyl-3-methyl-3-methyl-3-methyl-3-methyl-3-methyl-3-methyl-3-methyl-3-methyl-3-methyl-3-methyl-3-methyl-3-methyl-3-methyl-3-methyl-3-methyl-3-methyl-3-methyl-3-methyl-3-methyl-3-methyl-3-methyl-3-methyl-3-methyl-3-methyl-3-methyl-3-methyl-3-methyl-3-methyl-3-methyl-3-methyl-3-methyl-3-methyl-3-methyl-3-methyl-3-methyl-3-methyl-3-methyl-3-methyl-3-methyl-3-methyl-3-methyl-3-methyl-3-methyl-3-methyl-3-methyl-3-methyl-3-methyl-3-methyl-3-methyl-methyl-3-methyl-3-methyl-3-methyl-methyl-3-methyl-methyl-3-methyl-methyl-3-methyl-3-methyl-methyl-3-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-

The test chemical is then applied to the test chemical.

The test chemical is used to determine the concentration of the test substance in the test medium.

14 mg (0.025 mmol) of this intermediate was absorbed into 5 ml of dichloromethane, added to 1 ml of trifluoroacetic acid and stirred for 20 min at RT. The resulting 14 mg (98% dT) of the title compound was obtained by vacuum condensation and freeze-drying of the remaining dioxane residue.

The test chemical is then applied to the test chemical.

The test chemical is used to determine the concentration of the test substance in the test medium.

Intermediate 12 It consists predominantly of hydrocarbons having carbon numbers predominantly in the range of C1 through C4.]

The mixture was stirred for 24 h at RT. Then the same amount of 1,2-Oxopyrrolidine-1-yl-N- ((tert. butoxycarbonyl) L-tryptophanate was added again. After further stirring for 4 h, the solution was vacuum-dried, the residue was absorbed in a thick liquid and first diluted with sodium hydroxide dihydroxyethylamate (base compound 5) and then shaken with water. The residue was then washed with the corresponding fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorinated fluorin

The test chemical is then applied to the test chemical.

The test chemical is used to determine the concentration of the test substance in the test medium.

63 mg (0.167 mmol) of this intermediate was absorbed into 15 ml of dichloromethane, added to 2 ml of trifluoroacetic acid and treated in an ultrasonic bath for 40 min. The solution was then narrowed in vacuum and the dioxane/water residue lyophilized to obtain 65 mg (90% dT) of the title compound.

The test chemical is then applied to the test chemical.

The test chemical is used to determine the concentration of the test substance in the test medium.

Intermediate 13 It consists predominantly of hydrocarbons having carbon numbers predominantly in the range of C1 through C5 and boiling in the range of approximately -15oC to -15oC (-70oF to -15oF).]

The title compound was prepared by analogy to the synthesis of intermediate 7 by two steps from the dicyclohexylamine salt of parent compound 1 and (2S)-2-amino-3-(1H-indol-3-yl)-1-(1,2-oxazinane-2-yl) propane-1-on trifluoroacetic acid salt (intermediate 12).

The yield over two stages: 62 mg (67% d.i.)

The test chemical is then applied to the test chemical.

The test chemical is used to determine the concentration of the test substance in the test medium.

Intermediate 14 It consists predominantly of hydrocarbons having carbon numbers predominantly in the range of C1 through C5.]

The title compound was produced by analogy to the synthesis of intermediates 5 and 6 via two steps of N- (tert-butoxycarbonyl) L-phenylalanine and 1,2-oxazolidine hydrochloride (base compound 4).

The yield over two stages: 1650 mg (97% d.i.)

The test chemical is then applied to the test chemical.

The test chemical is used to determine the concentration of the test substance in the test medium.

Intermediate 15 It consists predominantly of hydrocarbons having carbon numbers predominantly in the range of C1 through C5.]

The title compound was prepared by analogy to the synthesis of intermediate 7 by two steps from the dicyclohexylamine salt of parent compound 1 and (2S)-2-anueno-1-(1,2-oxazolidin-2-yl) -3-phenylpropan-1-one trifluoroacetic acid salt (intermediate 14).

The yield over two stages: 110 mg (97% d.i.)

The test chemical is then applied to the test chemical.

The test chemical is used to determine the concentration of the test substance in the test medium.

Intermediate 16 It consists predominantly of hydrocarbons having carbon numbers predominantly in the range of C1 through C5.]

The title compound was produced in two steps from (βS) -N- (III-butoxycarbonyl) -β-methyl-L-phenylalanine and 1,2-oxazinane hydrochloride (parent compound 5) by analogy with the synthesis of intermediates 5 and 6.

The yield over two stages: 652 mg (63% d.o.h.)

The test chemical is then applied to the test chemical.

The test chemical is used to determine the concentration of the test substance in the test medium.

Intermediate 17 It consists predominantly of hydrocarbons having carbon numbers predominantly in the range of C1 through C5.]

The title compound was prepared by analogy to the synthesis of intermediate 7 by two steps from the dicyclohexylamine salt of parent compound 1 and (2S,3S)-2-amino-1-(1,2-oxazinan-2-yl)-3-phenylbutan-1-one trifluoroacetic acid salt (intermediate 16).

The yield over two stages: 101 mg (90% d.j.)

The test chemical is then applied to the test chemical.

The test chemical is used to determine the concentration of the test substance in the test medium.

Intermediate 18 It consists predominantly of hydrocarbons having carbon numbers predominantly in the range of C1 through C5.]

The title compound was produced by analogy to the synthesis of intermediates 5 and 6 via two steps from N- (tert-butoxycarbonyl) -3-nitro-L-tyrosine and 1,2-oxazine hydrochloride (starting compound 5).

The yield over 2 stages is 374 mg (47% of the dry weight)

The test chemical is then applied to the test chemical.

The test chemical is used to determine the concentration of the test substance in the test medium.

Intermediate 19 It consists predominantly of hydroxide, hydroxybenzene and hydroxybenzene compounds having carbon numbers predominantly in the range of C1 through C4.]

The title compound was produced by analogy with the synthesis of intermediate 7 by two steps from the dicyclohexylamine salt of parent compound 1 and (2S)-2-amino-3- ((4-hydroxy-3-nitrophenyl) -1,2-oxazinane-2-yl) propane-1-on trifluoroacetic acid salt (intermediate 18), and the product obtained at a purity of 63% was used as such in subsequent reactions without further purification.

The yield over two stages: 128 mg (61% d.j.)

The test chemical is then applied to the test chemical.

The test chemical is used to determine the concentration of the test substance in the test medium.

Intermediate 20 N- (tert-butoxycarbonyl) -N-methyl-L-valyl-N- (t) -R,3S,4S) -1-carboxy-2-methoxy-4-methylhexane-3-yl) -N-methyl-L-valinamide

51 mg (0.08 mmol) N-[(9H-fluorescent-9-ylmethoxy) carbonyl]-N-methyl-L-valyl-N-[(2R,3S,4S) -carb-oxy-2-methoxy-4-methylhexane-3-yl]-N-methyl-L-valinamide (Intermediate 4) were dissolved in 10 ml DMF and added to 0.5 ml piperidine. After 10 min of agitation in RT, the solution was narrowed in vacuum and the residue was stirred with diethyl ether. The insoluble components were filtered and washed with diethyl ether more than once. The filter residue was then narrowed to 5 ml diethyl ether/water (1:1) and the solution was added to 1 ml pentanoxide/rock (1:1) by adding 11 daltonic acid to the solution. The total amount of diethyl ether was reduced to 113% by pH. The product was further treated with 34 mg diethyl ether (1-39 mmol/l). The product was added to a vacuum with diethyl ether and added to a solution of diethyl ether and was subsequently added to the solution by adding 1 ml pentanoxide/water (1:1) and the solution was added to a total of 11 daltonic acid (13% by pH). The resulting solution was reduced to a total of 34 mg diethyl ether (99 mmol. The product was treated with diethyl ether and added to a solution of diethyl ether and was added to a solution of diethyl ether and was added to a solution of diethyl ether (1-39 mg diethyl ether). The product was added to a second dose of 34 mg diethyl ether and was added to the solution of diethyl ether and was added to the solution of diethyl ether and was added to a solution of diethyl ether and was added to the solution of diethyl ether. The product was added to a solution of diethyl ether and was added to the solution of diethyl ether and was added to the solution. The product was added to the solution was added to the solution was added to a solution of diethyl ether and

The test chemical is then applied to the test chemical.

The test chemical is used to determine the concentration of the test substance in the test medium.

Intermediate 21 It consists predominantly of hydrocarbons having carbon numbers predominantly in the range of C1 through C5.]

The title compound was produced in three steps by analogy with the synthesis of intermediates 5 and 6 by coupling of commercially available N-ter-butoxycarbonyl-3-methyl-L-valine with 1,2-oxazinian hydrochloride (starting compound 5), then defusing with trifluoroacetic acid and re-coupling with starting compound 1.

The test chemical is then applied to the test chemical.

The test chemical is used to determine the concentration of the test substance in the test medium.

Intermediate 22 The active substance is a compound with a specific chemical activity of not more than 0,5% by weight and a purity of not more than 0,5% by weight.

The title compound was produced in three steps by coupling of commercially available N-ter-butoxycarbonyl-β-phenyl-L-phenylalanine with 1,2-oxazinian hydrochloride (starting compound 5), followed by trifluoroacetic acid defusion and re-coupling with starting compound 1 by means of a preparative HPLC purification.

The test chemical is then applied to the test chemical.

The test chemical is used to determine the concentration of the test substance in the test medium.

Intermediate 23 The active substance is a compound with a specific chemical activity of not more than 0,5% by weight and a purity of not more than 0,5% by weight.

The title compound was produced in three steps by coupling of commercially available (1S,2S) -1-[[[tert. butoxycarbonyl) amino]-2-phenylcyclopropanocarboxylic acid with 1,2-oxazinian hydrochloride (base compound 5), followed by deflowering with trifluoroacetic acid and re-coupling with base compound 1 and the final product was purified by preparative HPLC.

The test chemical is then applied to the test chemical.

The test chemical is used to determine the concentration of the test substance in the test medium.

Intermediate 24 The active substance is a compound with a specific chemical activity of not more than 0,5% by weight and a purity of not more than 0,5% by weight.

The title compound was produced in three steps by coupling of commercially available (1S,2R)-1-[[[tert. butoxycarbonyl) amino]-2-phenylcyclopropanocarboxylic acid with 1,2-oxazinian hydrochloride (base compound 5), followed by deflowering with trifluoroacetic acid and re-coupling with base compound 1 and the final product was purified by preparative HPLC.

The test chemical is then applied to the test chemical.

The test chemical is used to determine the concentration of the test substance in the test medium.

Intermediate 25 The following substances are to be classified in the same heading as the active substance:

After 60 h of agitation in RT, the approach was vacuum-tighted and the residue was absorbed in 100 ml of ethyl alcohol. The organic phase was then vacuum-tighted. The reaction and further separation of the two diasters was performed by flash chromatography using 98:2 silica/ methanol/ electrolyte, which corresponds to the density of the residue. The reactions were combined in the vacuum.

Diastereomer one:

yield: 154 mg [still contaminated by N- (III-Butoxycarbonyl) L-phenylalanine which was separated at the subsequent stage]

The test chemical is a mixture of two or more substances.

Diastereomer two:

The yield is 86 mg (16% by weight)

The test chemical is a mixture of two or more substances.

Intermediate 26 The following substances are to be classified in the same heading as the active substance:

75 mg (approximately 0.22 mmol) of diastereomer 1 of intermediate 25 was absorbed into 10 ml of acetone and added to 228 mg (1.34 mmol) benzyl bromide, 616 mg (4.46 mmol) potassium carbonate and a spatula tip of tetra-n-butylammonium iodide. The solution was heated under reflux for 20 h and then pressed in vacuum. The residue was stirred with 100 ml dichloromethane, filtered and then pressed down the dichloromethane phase. The remaining residue was purified by preparative HPLC. 68 mg (72% d. th.) of the title compound was obtained.

The test chemical is then applied to the test chemical.

The test chemical is used to determine the concentration of the test substance in the test medium.

Intermediate 27 The following substances are to be classified in the same heading as the product:

83 mg (0.25 mmol) of diastereomer 2 of intermediate 25 was absorbed in 15 ml of acetone and mixed with 127 mg (0.74 mmol) benzyl bromide, 682 mg (4.93 mmol) potassium carbonate and a spatula tip of tetra-n-butylammonium iodide. The solution was heated under reflux for 20 h and then vacuum-pressed. The residue was mixed with 100 ml dichloromethane, filtered and then pressed down the dichloromethane phase. The remaining residue was purified by preparative HPLC. 87 mg (83% d. th.) of the title compound was obtained.

The test chemical is then applied to the test chemical.

The test chemical is used to determine the concentration of the test substance in the test medium.

Intermediate 28 It consists predominantly of hydrocarbons having carbon numbers predominantly in the range of C1 through C5.]

68 mg (0.16 mmol) tert-butyl-{(2S)-1-[[4-(benzyloxy) -1,2-oxazolidin-2-yl]-1-oxo-3-phenylpropan-2-yl}carbamates (diastereomer 1, intermediate 26) were incorporated into 10 ml of dichloromethane, added to 1 ml of trifluoroacetic acid and stirred for 30 min in RT.

The test chemical is then applied to the test chemical.

The test chemical is used to determine the concentration of the test substance in the test medium.

Intermediate 29 It consists predominantly of hydrocarbons having carbon numbers predominantly in the range of C1 through C5.]

82 mg (0.19 mmol) tert-butyl-{(2S)-1-[[4-(benzyloxy) -1,2-oxazolidin-2-yl]-1-oxo-3-phenylpropan-2-yl}carbamates (diastereomer 2, intermediate 27) were incorporated into 10 ml of dichloromethane, added to 1 ml of trifluoroacetic acid and stirred for 30 min in RT.

The test chemical is then applied to the test chemical.

The test chemical is used to determine the concentration of the test substance in the test medium.

Intermediate 30 The substance is to be classified in the immediate vicinity of the product and the product must be classified in the immediate vicinity of the product.

315 mg (0.494 mmol) N-[(9H-fluorescent-ylmethoxy) carbonyl]-N-methyl-L-valyl-N-[(2R,3S,4S) -1-carboxy-2-methyl-L-valyl-hexane-4-methyl-3-yl]-N-methyl-L-valinamide (Intermediate 4) were dissolved in 12 ml DMF, with 104 mg (0.543 mmol) 1-dimethylamylamypropyl) 3-ethylcarbodimide hydrochloride and 83 mg (0.543 mmol) 1-hydroxy-1H-benzotriazole hydrate dissolved unprotected and 90 min at RT Boost. Subsequently 112 μl N-methyl-diisoproplamine and 149 mg (2,449 mmol) high-pressure (2,3R-3-3-methyl-methyl-methyl-3-pyrol) were dissolved in a vacuum-formed solution containing 140 mg T-methyl-pyrol (35%). The second group of preparations was obtained by adding 1 mg of T-propyl-acetyl-acetyl to the solution and leaving it in the vacuum-formed form of a second solution, which was then released by the second dose of the drug.

The test chemical is then applied to the test chemical.

The test chemical is used to determine the concentration of the test substance in the test medium.

Intermediate 31 [[[1S,2R)-1-Amino-2-phenylcyclopropyl]][[1,4-dihydro-3H-2,3-benzoxazine-3-yl) methane trifluoroacetic acid salt

The title compound was prepared by analogy to the synthesis of intermediate 10 from tert-butyl[(1S,2R)-1- ((hydroxycarbamoyl)-2-phenylcyclopropyl]carbamate (primary compound 8).

The test chemical is then applied to the test chemical.

The test chemical is used to determine the concentration of the test substance in the test medium.

Intermediate 32 The following substances are to be classified in the same heading as the active substance:

The organic phase was dried, filtered and compacted using magmus sulphate. The residue was collected in 16 ml DMF and successively purified with 365 mg (1 mmol) tert-butyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-meth

The test chemical is then applied to the test chemical.

283 mg (0.466 mmol) of this intermediate was absorbed into 5 ml of dichloromethane, mixed with 5 ml of anhydrous trifluoric acid and stirred for 2 h at RT. The solution was then narrowed in a high vacuum and the remaining residue was cleaned by HPLC preparation to give 156 mg (61% d.i.t.) of the title compound as a colourless foam.

The test chemical is then applied to the test chemical.

The test chemical is used to determine the concentration of the test substance in the test medium.

Other (2R)-2-Amino-1- ((1,2-Oxazinan-2-yl) 3-phenylpropan-1-one salt of trifluoroacetic acid

The title compound was produced by analogy to the synthesis of intermediate 6 from N- (tert-butoxycarbonyl) D-phenylalanine.

The test chemical is then applied to the test chemical.

The test chemical is used to determine the concentration of the test substance in the test medium.

Intermediate 34 It consists predominantly of hydrocarbons having carbon numbers predominantly in the range of C1 through C5.]

The title compound was produced by analogy to the synthesis of intermediate 6 from commercially available N- (tert-butoxycarbonyl) -α-methyl-L-phenylalanine.

The test chemical is then applied to the test chemical.

The test chemical is used to determine the concentration of the test substance in the test medium.

Examples of implementations: Example 1 It consists predominantly of hydrocarbons having carbon numbers predominantly in the range of C1 through C5 and boiling in the range of approximately -15oC to -15oC (-70oF to -15oF).]

143 mg (0.223 mmol) N-[(9H-fluorene-9-ylmethoxy) carbonyl]-N-methyl-L-valyl-N-[((2R,3S,4S) -N-carboxy-2-methyl-L-valyl-N-methylhexane-3-yl]-N-methyl-L-valinamide (Intermediate 4) were absorbed in 15 ml DMF and subsequently with 141 mg (0.22 mmol) (2R,3R) -Methoxy-2-methyl-N-[(2S)-1-(1,2-oxazin-2-methyl) -N-dioxophenyl-L-valyl-N-valyl]-3-[(2R,3S) -Pyrroxy-2-methyl]propanidyl-methyl-acetyl-acetyl (Intermediate 7), 102 mg (0.27 mg) Oxazin-3-L-methyl-L-carboxyl-N-nitrogen-3-N-dioxophenol was removed from the sample, and the solution was mixed with a mixture of N-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-meth

The test chemical is then applied to the test chemical.

The test chemical is used to determine the concentration of the test substance in the test medium.

After adding 1 ml of piperidine, the solution was stirred for 1 h at RT, then vacuum-tighted and the residue was cleaned by HPLC preparation (eluent: acetonitrile + 0.01% TFA/ water + 0.01% TFA). 22 mg (54% dT) of the titration was obtained as a colourless foam.

The test chemical is then applied to the test chemical.

The test chemical is then applied to the test chemical and the test chemical is then removed.

The mean values for the H-NMR (600 MHz, DMSO-d6): δ = 8.8 (m, 2H), 8.7 (m, 1H), 8.42 and 8.15 (2d, 1H), 7.3-7.1 (m, 5H), 5.12 and 4.95 (2m, 1H), 4.70 and 4.62 (2m, 1H), 4.62 and 4.50 (2t, 1H), 4.1 to 3.9 (m, 3H), 3.85 (m, 1H), 3.75 to 3.6 (m, 2H), 3.23, 3.18, 3.17, 3.14, 3.02 and 2.96 (6s, 9H), 3.1-2.9 and 2.75 (2m, 2H), 2.46 (m, 3H), 2.4-2.1 (m, 2H), 2.05 (m, 2H), 2.85 to 1.55 (m, 6H), 1.5-1.2 (m, 3H), 1.18 to 0.8 (m, 18H), 3H (H), 3H (H), 3H (H), 3H (H), 3H (H), 3H (H), 3H (H), 3H (H), 3H (H), 3H (H), 3H) are among the most widely used H-contacted signals.

Example 2 The substance is to be classified in the immediate vicinity of the product and the product must be classified in the immediate vicinity of the product.

50 mg (0.0411 mmol) N-Methyl-L-valyl-N-[(3R,4S,5S) -3-methoxy-1-[(2S)-2-[(1R,2R) -1-methoxy-2-methyl-3-{(2S)-1-(1,2-oxaziman-2-yl) -1-oxo-3-phenylpropan-2-yl]amino}-3-oxopropyl]pyrrolidin-1-yl}-5-methyl-1-oxoheptan-4-yl]-N-methyl-L-valynamide trifluoric acid salt (Example 1) were dissolved in 3 ml of dioxane/water (1:1) phase and then dissolved in 4.9 mg (0.08 mmol) of dimethyl-ethyl glycolide hydrochloride.

The test chemical is then applied to the test chemical.

The test chemical is then applied to the test chemical and the test chemical is then removed.

The mean values for the 1H-NMR (600 MHz, DMSO-d6): δ = 9.2 (m, 1H), 8.9 (m, 1H), 8.4 and 8.15 (2d, 1H), 7.3-7.1 (m, 5H), 5.12 and 4.95 (2m, 1H), 4.70 and 4.62 (2m, 1H), 4.62 and 4.55 (2t, 1H), 4.1-3.9 (m, 3H), 3.9-3.7 (m, 5H), 3.23, 3.18, 3.17, 3.15, 3.02 and 2.98 (6s, 9H), 2.95 and 2.75 (2m, 2H), 2.8 (m, 3H), 2.46-2.00 (m, 4H), 1.85-1.55 (m, 6H), 1.5-1.2 (m, 3H), 1.1-0.8 (m, 18H), 3.75 (t, 0.2H) [H2 Peak under extended HO-signal]

Example 3 The substance is to be classified in the immediate vicinity of the active substance and the active substance is to be classified in the active substance.

20 mg (0.022 mmol) N-Methyl-L-valyl-N-[(3R,4S,5S) -3-methoxy-1-{(2S)-2-[(1R,2R) -1-methoxy-2-methyl-3-{(2S)-1-(1,2-oxazinan-2-yl) -1-oxo-3-phenylpropan-2-yl]amino}-3-oxopropyl]pyrrolidine-1-yl}-5-methyl-1-oxoheptan-4-yl]-N-methyl-L-valinamide trifluoroacetic acid salt (Example 1) were dissolved in 1 ml DMF, transferred to 3.4 mg (1 μl) of iodomethan and 7.6 mg (0.055 ml) of potassium carbonate and stirred at 1 HP. Subsequently, the same amount of potassium carbonate was added to the bath and treated with a vacuum bath (4 mmol) and 8 mg of the residual solution was removed from the bath.

The test chemical is then applied to the test chemical.

The test chemical is then applied to the test chemical and the test chemical is then removed.

The mean value of the measurements of the effects of the test chemical on the human body is calculated by dividing the mean value of the measurements by the mean value of the measurements of the human body.

Example 4 The substance is to be classified in the immediate vicinity of the product and the product must be classified in the immediate vicinity of the product.

126 mg (0.198 mmol) N-[(9H-fluorescent-9-yhnethoxy) carbonyl]-N-methyl-L-valyl-N-[((2R,3S,4S) -N-carboxy-2-methyl-L-valyl-N-[(2R,3S,4S]-carboxy-2-methyl-Methoxy-4-methylhexane-3-yl]N-methyl-L-valinamide (Intermediate 4) were absorbed in 10 ml DMF and subsequently 105 mg (0.198 mmol) (2R,3R) -Methoxy-2-methyl-N-[(2S,3S)-1-(1,2-oxazine-2--2-methyl) -N-2-oxo-2-phenyl-Nesibutyl]-3-[(2-S) -N-2-methyl-methyl-2-methylhydroxy-2-methoxy) -propanoyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl

The test chemical is then applied to the test chemical.

The test chemical is used to determine the concentration of the test substance in the test medium.

After adding 1 ml of piperidine, the solution was stirred for 1 h at RT, then vacuum-tighted and the residue was cleaned by HPLC preparation (eluent: acetonitrile + 0.01% TFA/ water + 0.01% TFA). 91 mg (46% dT) of the titration was obtained as a colourless foam.

The test chemical is then applied to the test chemical.

The test chemical is then applied to the test chemical and the test chemical is then removed.

The mean values for the 1H-NMR (600 MHz, DMSO-d6): δ = 8.87 and 8.80 (2d, 2H), 8.75 (m, 1H), 8.40 and 7.98 (2d, 1H), 7.3-7.1 (m, 5H), 5.45 and 5.2 (2t, 1H), 4.78 and 4.62 (2m, 1H), 4.73 and 4.58 (2t, 1H), 4.2-4.0 (m, 3H), 3.7-3.6 (m, 1H), 3.35, 3.20, 3.18, 3.14, 3.12 and 3.00 (6s, 9H), 3.1 and 2.95 (2m, 2H), 2.46 (m, 3H), 2.4-2.0 (m, 4H), 1.9-1.6 (m, 4H), 1.6-1.2 (m, 5H), 1.1-0.75 (m, 21H), 0.80 (t, 3H) [further signals under H2O Peak]

Example 5 The substance is to be classified in the immediate vicinity of the product and the product must be classified in the immediate vicinity of the product.

138 mg (0.216 mmol) N-[(9H-fluorine-9-ylmethoxy) carbonyl]-N-methyl-L-valyl-N-[(((2R,3S,4S) -N-carboxy-2-methyl-L-valyl-N-[(2R,3S,4S) -N-carboxy-2-methyl-Hexane-4,5-methylhexane-3-yl]-N-methyl-L-valinamide (Intermediate 4) were ingested in 14 ml DMF and were successively administered with 109 mg (0.216 mmol) (2R,3R) -3-methyl-N-N-[[2S]-1-(1,2-oxazolidin-2-carbonyl) -1-dioxy-2-methyl-methyl-propanol]-3-[(2S) -2-pyrroxy-2-methyl-hydroxy-2-methyl-valyl]-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-

The test chemical is then applied to the test chemical.

The test chemical is used to determine the concentration of the test substance in the test medium.

After adding 1 ml of piperidine, the solution was stirred for 1 h at RT, then vacuum-tighted and the residue was cleaned by HPLC preparation (eluent: acetonitrile + 0.01% TFA/ water + 0.01% TFA). 37 mg (24% dT) of the titration was obtained as a colourless foam.

The test chemical is then applied to the test chemical.

The test chemical is then applied to the test chemical and the test chemical is then removed.

The mean values for the 1H-NMR (600 MHz, DMSO-d6): δ = 8.9-8.7 (m, 3H), 8.5 and 8.2 (2m, 1H), 7.3-7.1 (m, 5H), 5.6 and 5.4 (2m, 1H), 4.7 and 4.6 (2m, 1H), 4.65 and 4.55 (2t, 1H), 4.0-3.9 (m, 3H), 3.75-3.6 (m, 2H), 3.23, 3.20, 3.15, 3.05 and 2.98 (5s, 9H), 3.0 and 2.7 (2m, 2H), 2.46 (m, 3H), 2.35-2.15 (m, 4H), 2.1-2.0 (m, 2H), 1.85-1.6 (m, 3H), 1.45 (m, 1H), 1.25 (m, 1H), 1.1-0.85 (m, 18H), 1875 (t, 3H) [H2] are further described in H2O Peak.

Example 6

The substance is to be classified in the immediate vicinity of the product and the product must be classified in the immediate vicinity of the product.

44.5 mg (0.071 mmol) N- (((9H-fluoren-9-ylmethoxy) carbonyl) -N-methyl-L-valyl-N- (((2R,3S,4S) -N-carboxy-2-methyl-L-valyl-N-methylhexane-3-yl) -N-methyl-L-valinamide (Intermediate 4) were ingested in 10 ml DMF and then successively with 38.6 mg (0.071 mmol) (2R,3R) -N-methyl-N-N-Fluoren-9-yl) -N-carbonyl) -N- ((9H-fluoren-9-ylmethoxy) carbonyl) -N-methyl-L-valyl-N-valyl-N-valyl-N-N-[(2R,3S,4S) -N-methyl-N-methyl-L-valyl-L-valyl-N-valyl-N-N- (Intermediate 4) ], followed by 32.5 mg (0.86 mmol) (38.6 mg (0.071 mmol) (38.6 mg) (38.0 mmol) (38.0 mmol)), followed by 41 mg (0.0 mmol) (92.0 mmol) (32.0 mmol) (32.0 mmol) (32.0 mmol) (32.0 mmol) (32.0 mmol) (32.0 mmol) (32.0 mmol) (32.0 mmol) (32.0 mmol) (32.0 mmol) (32.0 mmol) (32.0 mmol) (32.0 mmol) (32.0 mmol) (32.0 mmol) (32.0 mmol) (32.0 mmol) (32.0 mmol) (32.0 mmol) (32.0 mmol) (32.0 mmol) (32.0 mmol) (31.0 mmol) (31.0 mmol) (31.0 mmol) (31.8 mmol) (31.8 mmol) (31.8 mmol) (31.8 mmol) (31.8 mmol) (31.3) (31.3) (31) (31) (31) (31)) and 0.0 mmol) The result was obtained by replacing the organic carboxyl-methyl-methyl-meth

The test chemical is then applied to the test chemical.

The test chemical is used to determine the concentration of the test substance in the test medium.

After decanting the ether, the residue was cleaned by HPLC preparation (eluent: acetonitrile / 0.1% aq. TFA) and 16 mg (26% d. th.) of the titer compound was obtained as a colourless foam.

The test chemical is then applied to the test chemical.

The test chemical is then applied to the test chemical and the test chemical is then removed.

The mean value of the 1H-NMR (400 MHz, DMSO-d6): δ = 8.9-8.6 (m, 3H), 8.4, 8.3, 8.1 and 8.0 (4d, 1H), 7.3-7.1 (m, 5H), 6.7-6.5 (m, 2H), 5.2-4.8 (m, 3H), 4.75-4.55 (m, 3H), 4.05-3.95 (m, 1H), 3.7-3.4 (m, 4H), 3.22, 3.17, 3.15, 3.02 and 2.95 (6s, 9H), 3.0 and 2.7 (2 br. m, 2H), 2.46 (m, 3H), 2.4-1.2 (br. m, 13H), 1.1-0.85 (m, 18H), 0.75 (m, 3H) [further signals are hidden under H2O-Peak].

Example 7 The substance is to be classified in the immediate vicinity of the product and the product must be classified in the immediate vicinity of the product.

N-[(9H-fluorene-9-ylmethoxy) carbonyl]-N-methyl-L-valyl-N-[((2R,3S,4S) -1-carboxy-2-methyl-methyl-methyl-hexane-3-yl]-N-methyl-L-valinamide (Intermediate 4) were absorbed in 13 ml DMF and subsequently filtered with 145 mg (0.251 mmol) (2R,3R) -N-(2S) -9H-fluorene-9-ylmethoxy) carbonyl-N-methyl-L-valyl-N-[(2R,3S,4S) -N-carboxy-2-methyl-methyl-hexane-3-methyl-N-carboxy-2-methyl-methyl-methyl-methyl-methyl-hexane-4-yl]-N-methyl-methyl-L-valinamide (Intermediate) in 13 ml DMF, then further filtered with 145 mg (0.251 mmol) (2R,3R) -N-(2S) -N-nitroproproproprophenol-N-N-nitrophenyl-3-methyl-methyl) (2-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl) (1-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl)), then further filtered with 19 mg (0.27-36-56-5-3-3-3-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-

After adding 0.6 ml piperidine, the solution was stirred for 1 h at RT, then vacuum-tighted and the residue was cleaned by preparative HPLC (eluent: acetonitrile + 0.01% TFA/ water + 0.01% TFA). 105 mg (45% DT) of the titrate was obtained as a slightly yellowish foam.

The test chemical is then applied to the test chemical.

The test chemical is then applied to the test chemical and the test chemical is then removed.

The mean values of the H-NMR (500 MHz, DMSO-d6) are: δ = 10.85 and 10.72 (2s, 1H), 8.9-8.6 (m, 3H), 8.45 and 8.2 (2d, 1H), 7.70 and 7.72 (2s, 1H), 7.40 and 7.32 (2d, 1H), 7.01 and 7.00 (2d, 1H), 5.0 to 4.85 (m, 1H), 4.15 to 4.0 (m, 2H), 3.95 to 3.75 (m, 2H), 3.7-3.6 (m, 2H), 3.28, 3.21, 3.16, 3.15, 3.02 and 2.96 (6m, 9H), 2.9 and 2.75 (2m, 2H), 2.46 (m, 3H), 2.4-2.3 (m, 2H), 2.3-2.2 (m, 1H), 2.95 to 1.1-1, 1.85 to 1.1-1, 1.85 to 1.2-1, 1.75 to 1.5-1, 1.75 to 1.5-1, 1.75 to 1.5, 1.75 to 1.75 (m, 2H), 1.75 to 1.75 to 1.75 (m, 1H), 1.75 to 1.75 to 1.75 (m, 1H), 1.75 to 1.75 to 1.75 (m, 1H), 1.75 to 1.75 to 1.75 (m, 1H), 1.75 to 1.75 to 1.75 to 1.75 (m, 1H, 1H, 2H, 1H, 1H, 1H, 2H, 1H, 1H, 2H, 1H, 1H, 2H, 1H, 2H, 1H, 2H, 1H, 2H, 1H, 2H, 2H, 1H, 2H, 1H, 2H, 2H, 2H, 1H, 2H, 2H, 2H, 2H, 1H, 2H, 2H, 2H, 2H, 2H, 2H, 2H, 2H, 2H, 2H, 2H, 2H, 2H, 2H, 2H, 2H, 2H, 2H, 2H, 2H, 2H, 2H, 2H, 2H, 2H, 2H, 2H, 2H, 2H, 2H, 2H, 2H, 2H, 2H, 2H, 2

Example 8 The substance is to be classified in the immediate vicinity of the product and the product must be classified in the immediate vicinity of the product.

15.8 mg (0.025 mmol) N-[(9H-fluorescent-ylmethoxy) carbonyl]-N-methyl-L-valyl-N-[(2R,3S,4S) -1-carboxy-2-methyl-methoxy-4-methylhexane-3-yl]-N-methyl-L-valinamide (Intermediate 4) were ingested in 5 ml DMF and were followed up with 14 mg (0.025 mmol) (2R,3R) -N-[(2S) -1,4-dihydroxy-3H-2,3-benzoxazine-3-di-methyl) -1-oxo-3-phenylpropanyl-2-methyl-2-methyl-2-methyl-3-methoxy-2-methyl-methoxy-2-methyl-methoxy-3-[S]-pyrroxy-2-methylpropanyl-2-methyl]-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-hexane-3-hexane-4-yl (Intermediate) 11 mg (0.03 mmol) and then replaced by 11 mg (0.03 mmol) of Oxy-3-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl

The test chemical is then applied to the test chemical.

The test chemical is used to determine the concentration of the test substance in the test medium.

After adding 0.5 ml piperidine, the solution was stirred for 10 min at RT, then vacuum-tighted and the residue was cleaned by preparative HPLC (eluent: acetonitrile / 0.1% aq. TFA) to obtain 10 mg (97% d. th.) of the titer compound.

The test chemical is then applied to the test chemical.

The test chemical is then applied to the test chemical and the test chemical is then removed.

The mean value of the 1H-NMR (400 MHz, DMSO-d6): δ = 8.90-8.62 (m, 3H), 8.60 and 8.33 (2d, 1H), 7.35-7.1 (m, 9H), 5.4-5.0 (m, 4H), 4.7-4.5 (m, 3H), 3.95 (m, 1H), 3.7-3.4 (m, 3H), 3.24, 3.20, 3.18, 3.16, 3.10 and 3.08 (6s, 9H), 3.0 and 2.85 (2 br. m, 2H), 2.46 (m, 3H), 2.3 (br. m, 2H), 2.05 (br. m, 2H), 1.9-1.6 (m, 3H), 1.5-1.2 (br. m, 2H), 1.1-0.85 (m, 18H), 0.75 (m, 3H) [further H2O-Peak signals are hidden].

Example 9 N-methyl-L-valyl-N-[(3R,4S,5S)-1-{(2S)-2-[(1R,2R)-3-{(2S)-3-(1H-indol-3-yl)-1-(1,2-oxazinane-2-yl)-1-oxopropan-2-yl]amino}-1-methoxy-2-methyl-3-oxopropyl]pyrrolidine-1-yl}-3-methoxy-5-methyl-1-oxoheptan-4-yl]-N-methyl-L-valinamide Trifluoric acid salt

40 mg (0.076 mmol) N-methyl-L-valyl-N- (2R,3S,4S) -1-carboxy-2-methyl-oxy-4-methylhexan-3-yl) -N-methyl-L-valinamide (intermediate 20) were absorbed in 5 ml DMF and were followed by 43 mg (0.078 mmol) (2R,3R) -N- (2S) -3-(2S) -Indol-3-propyl) -1,2-oxazin-2-yl) -1-oxopropan-2-diyl]-3-methyl-methyl-2-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl

The test chemical is then applied to the test chemical.

The test chemical is used to determine the concentration of the test substance in the test medium.

After adding 0.5 ml of trifluoroacetic acid, the solution was treated in the ultrasonic bath for 30 min and then stirred for another 30 min in RT, then vacuum-pressed, the residue was added with acetonitrile and re-pressed.

The test chemical is then applied to the test chemical.

The test chemical is then applied to the test chemical and the test chemical is then removed.

The mean values for the 1H-NMR (400 MHz, DMSO-d6): δ = 10.8 and 8.8 (2m, 3H), 8.35 and 8.05 (2d, 1H), 7.55 (m, 1H), 7.3 (d, 1H), 7.20 to 6.91 (m, 3H), 5.12 and 4.95 (2m, 1H), 4.7-4.5 (m, 2H), 4.1-3.9 (m, 2H), 3.85 (m, 2H), 3.75 to 3.4 (m, 5H), 3.21, 3.15, 3.14, 3.10, 2.95 and 2.85 (6s, 9H), 2.46 (m, 3H), 2.4-2.2 (m, 2H), 2.1-1.9 (m, 2H), 1.85 to 1.65 (m, 4H), 1.65 to 1.2 (m, 3H), 1.05 to 1.0d, 3H), 0.8 to 0.95 (m, 2H), 0.75 to 3M (H), 3H, H2 are further described in H-Peak [H2].

Example 10 The substance is to be classified in the immediate vicinity of the product and the product must be classified in the immediate vicinity of the product.

25 mg (0.026 mmol) N-methyl-L-valyl-N-[(3R,4S,5S)-1-{(2S)-2-[(1R,2R)-3-{[(2S)-3-(4-hydroxy-3-nitrophenyl)-1-(1,2-oxazinane-2-yl)-1-oxopropan-2-yl]amino}-1-methoxy-2-methyl-3-oxopropyl]-pyrrolidine-1-yl}-3-methoxy-5-methyl-1-oxoheptane-4-yl]-N-methyl-L-valine trifluoric acid saline (Example 7) were dissolved in 2 ml 1 N of hydrochloric acid and then lyophilised, resulting in 16.41% (7 d. mg of the active substance) being readily soluble as a saline solution.

The test chemical is then applied to the test chemical.

The test chemical is used to determine the concentration of the test substance in the test medium.

Example 11 N-methyl-L-valyl-N-[(3R,4S,5S)-1-{(2S)-2-[(1R,2R)-3-(({(2S)-1-[4-(benzyloxy)-1,2-oxazolidin-2-yl]-1-oxo-3-phenylpropan-2-yl) amino)-1-methoxy-2-methyl-3-oxopropyl]pyrrolidin-1-yl}-3-methyl-oxy-5-methyl-1-oxoheptan-4-yl]-N-methyl-L-valinamide Trifluoroacetic acid salt (diastereomer 1)

The title compound was produced by coupling of parent compound 1 with intermediate 28, then dilution with Trifluoroesic acid, then coupling with intermediate 4 and finally dilution with piperidine, in analogy to the synthesis in example 8, and purification of the final product by preparative HPLC (eluent: acetonitrile / 0.1% aq. TFA) resulting in 16.4 mg of title compound.

The test chemical is then applied to the test chemical.

The test chemical is then applied to the test chemical and the test chemical is then removed.

The mean value of the dose of the active substance is calculated by dividing the dose by the mean value of the dose of the active substance in the dose range from 1 hour to 1 hour.

Example 12 N-methyl-L-valyl-N-[(3R,4S,5S)-1-{(2S)-2-[(1R,2R)-3-(({(2S)-1-[4-(benzyloxy)-1,2-oxazolidin-2-yl]-1-oxo-3-phenylpropan-2-yl) amino)-1-methoxy-2-methyl-3-oxopropyl]pyrrolidin-1-yl}-3-methyl-oxy-5-methyl-1-oxoheptan-4-yl]-N-methyl-L-valinamide Trifluoroacetic acid salt (diastereomer 2)

The title compound was produced by coupling of parent compound 1 with intermediate 29, then dilution with Trifluoroesic acid, then coupling with intermediate 4 and finally dilution with piperidine, in analogy to the synthesis in example 8, and purification of the final product by preparative HPLC (eluent: acetonitrile / 0.1% aq. TFA).

The test chemical is then applied to the test chemical.

The test chemical is then applied to the test chemical and the test chemical is then removed.

The mean value of the dose of the active substance is calculated by dividing the dose by the mean value of the dose of the active substance in the dose range from 1 hour to 1 hour.

Example 13 The following substances are to be classified in the same category as the active substance:

27 mg (0.03 mmol) of the compound from example 12 were taken in a mixture of 42 ml THF and 22 ml water, diluted with 1.35 ml 2 M hydrochloric acid and hydrated for 20 min at RT and normal pressure over 10% palladium/carbon. The catalyst was then de-filtered and the reaction mixture neutralized with aqueous sodium hydrocarbonate solution. Then extracted with ethyl acetate. After separation of the phases, the ethylacination phase was narrowed and the residue was purified by flash chromatography on a silica gel with dichloromethane/methanol/17% aq. ammonia (125:3:0.3). The corresponding reactions were eased and the residue was removed from the diisoxide.

The test chemical is then applied to the test chemical.

The test chemical is then applied to the test chemical and the test chemical is then removed.

The mean value of the dose of the active substance is calculated by dividing the dose by the mean value of the dose of the active substance, which is calculated by dividing the dose by the mean value of the dose of the active substance by the mean value of the dose of the active substance.

Example 14 N-methyl-L-valyl-N-[(3R,4S,5S)-1-{(2S)-2-[(1R,2R)-3-{(2S)-3,3-dimethyl-1- ((1,2-oxazinane-2-yl)-1-oxobutan-2-yl]amino}-1-methoxy-2-methyl-3-oxopropyl]pyrrolidine-1-yl}-3-methoxy-5-methyl-1-oxoheptan-4-yl]-N-methyl-L-valinamide Trifluoroacetic acid salt

The final product was purified by preparative HPLC (elute: acetonitrile + 0.01% TFA/ water + 0.01% TFA) and 2.5 mg of the final product was obtained.

The test chemical is then applied to the test chemical.

The test chemical is used to determine the concentration of the test substance in the test medium.

Example 15 N-methyl-L-valyl-N-[(3R,4S,5S) -3-methoxy-1-{(2S)-2-[(1R,2R) -1-methoxy-2-methyl-3-{(2S)-1-(1,2-oxazinan-2-yl) -1-oxo-3,3-diphenylpropan-2-yl]amino}-3-oxopropyl]pyrrolidin-1-yl}-5-methyl-1-oxoheptan-4-yl]-N-methyl-L-valinamide Trifluoroacetic acid salt

The title compound was prepared in three steps from intermediate 22 by first dilution with Trifluoroacetic Acid, then coupling with intermediate 4 and finally dilution with Piperidine.

The test chemical is then applied to the test chemical.

The test chemical is used to determine the concentration of the test substance in the test medium.

Example 16 The substance is to be classified in the immediate vicinity of the product and the product must be classified in the immediate vicinity of the product.

The title compound was prepared in three steps from intermediate 23 by first dilution with Trifluoroacetic Acid, then coupling with intermediate 4 and finally dilution with Piperidine.

The test chemical is then applied to the test chemical.

The test chemical is used to determine the concentration of the test substance in the test medium.

The mean value of the dose of the active substance is calculated by dividing the dose by the mean value of the dose of the active substance, which is calculated by dividing the dose by the mean value of the dose of the active substance by the mean value of the dose of the active substance.

Example 17 The substance is to be classified in the immediate vicinity of the product and the product must be classified in the immediate vicinity of the product.

617 mg (1.2 mmol) tert-butyl- ((2S)-2-[(1R,2R)-1-methoxy-2-methyl-3-{[(1S,2R)-1-(1,2-oxazinane-2-ylcarbonyl)-2-phenylcyclopropyl]amino}-3-oxopropyl]pyrrolidine-1-carboxylate (intermediate 24) was taken in 44 ml of dichloromethane, added to 4.4 ml of trifluoroacetic acid and stirred for 30 min. The remaining residue was then vacuumed and lyophilised from dioxane/water. 702 (quant.) mg (2R,3R)-3-methoxy-2-methyl-N[(1S,2R)-1-,3-oxazinane-2-carbonyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-

470 mg (0.74 mmol) N-[(9H-fluorescent-9-ylmethoxy) carbonyl]-N-methyl-L-valyl-N-[((2R,3S,4S) -1-carboxy-2-methyl-4-methylhexane-3-yl]-N-methyl-L-valinamide (Intermediate 4) were absorbed into 57 ml DMF and subsequently reconstituted with 390 mg (approximately 0.74 mmol) of the above (2R,3R) -3-methyl-N-methyl-N-carbonyl) N-[(9H-fluorescent-9-ylmethoxy) carbonyl-N-[(1S,2R) -1,2-dioxyn-2-methyl-L-valyl-L-valyn-2-methyl-N-cycloprophenyl]-3-methyl-N-[(2S,4S) -pyrroxydehydroxy-2-carbonyl] Anadrol-methyl-acetyl-fluoric acid, 336 mg (0.88-L-methyl-N-methyl-L-methyl-N-methyl-N-methyl-N-methyl-N-oxy-acetyl) was dissolved into 390 mg (approximately 0.74 mmol) of the above-mentioned (2R,3R-3-R) -3-methyl-N-N-methyl-N-N-N-methyl-N-N-methyl-N-N-N-methyl-N-N-N-methyl-N-N-N-O2-methyl-N-O2-methyl-N-N-O2-methyl-N-N-O2-methyl-N-N-N-O2-2-2-methyl-N-N-N-O2-methyl-N-N-N-N-O2-N-O2-N-O2-N-N-O2-N-N-N-N-N-O2-N-N-N-N-N-N-N-N-N-O2-N-N-N-N-N-N-N-N-N-N-N-N-N-N-N-

The test chemical is then applied to the test chemical.

The test chemical is used to determine the concentration of the test substance in the test medium.

After adding 2.4 ml piperidine, the solution was stirred for 30 min at RT, then vacuum-tighted and the residue was cleaned by preparative HPLC (eluent: acetonitrile / 0.1% aq. TFA). 260 mg (64% d.i.t.) of the titer compound was obtained as a colourless foam.

The test chemical is then applied to the test chemical.

The test chemical is then applied to the test chemical and the test chemical is then removed.

The mean value of the measurements of the test chemical is calculated by dividing the mean value of the measurements of the test chemical by the mean value of the measurements of the test chemical.

Example 18 The substance is to be classified in the immediate vicinity of the product and the product must be classified in the immediate vicinity of the product.

166 mg (0.196 mmol) N-[9H-fluorene-9-ylmethoxy) carbonyl-valinamide (intermediate 30) were absorbed in 40 ml DMF and were followed by 80 mg (0.196 mmol) [3R,4S,5S) N-methyl-L-valyl-N-methyl-1-methyl]1,4-dibenzoyl-3-valyl-3-methoxy-1-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-meth

The test chemical is then applied to the test chemical.

The test chemical is used to determine the concentration of the test substance in the test medium.

After adding 817 μl piperidine, the solution was stirred for 5 min at RT, then compressed in a vacuum and the residue was first digested with diethyl ether and then cleaned by HPLC (eluent: acetonitrile + 0.1% TFA / 0.1% aq. TFA). The corresponding fractions were combined, the solvent removed in a vacuum and the residue then lyophilised from dioxane/water. 12 mg (92% d. th.) of the titration compound was obtained as a colourless foam.

The test chemical is then applied to the test chemical.

The test chemical is used to determine the concentration of the test substance in the test medium.

Example 19 The substance is to be classified in the immediate vicinity of the product and the product must be classified in the immediate vicinity of the product.

100 mg (0.181 mmol) N-[Benzyloxy) carbonyl]-N-methyl-L-threonyl-N-[((2R,3S,4S) -carboxy-2-methoxy-4-methylhexane-3-methyl]N-methyl-L-valinamide (Intermediate 32) were absorbed in 5 ml DMF and were followed by 94 mg (0.181 mmol) (2R,3R) -3-methoxy-2-methyl-N-((2S) -1-oxazine-2-an-2-methyl) -1-oxo-2-phenylpropan-2-diol]-3-[[2S]-pyrrolidin-2-methyl]propan-2-carboxy-2-methoxy-hydroxy acid (Intermediate 7), 42 mg (0.218,5 mmol) 1-dimethyl-3-methoxy-3-benzoyl-benzoyl-benzoyl-benzoyl-benzoyl (0.2Mg) The residue was removed from the vacuum and the residue was then removed by the retention of the N-methyl-methoxy-1-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-methyl-meth

The test chemical is used to determine the concentration of the test substance in the test medium.

100 mg (0.107 mmol) of this intermediate was dissolved in 20 ml of methanol and hydrated at RT and normal pressure for 1 h over 10% palladium/carbon. The catalyst was then filtered and the solvent evaporated. The remaining residue was lyophilised from dioxane/water (1:1) to obtain 88 mg (97% dT) of the title compound.

The test chemical is used to determine the concentration of the test substance in the test medium.

Example 20 The following substances are to be classified in the same category as the active substance:

84 mg (0.105 mmol) N-Methyl-L-threonyl-N-[(3R,4S,5S) -3-methoxy-1-{(2S)-2-[(1R,2R) -1-meth-oxy-2-methyl-3-{(2S)-1-(1,2-oxazinan-2-yl) -1-oxo-3-phenylpropan-2-yl]amino-3-oxopropyl]-pyrrolidin-1-yl}-5-methyl-1-oxoheptan-4-yl]-N-methyl-L-valinamide (Sample 19) were obtained in 5 ml of dioxane/water (1:1) in the dry phase and 31 μl (0.418 mmol) of 37% aqueous hydrocarbon foracid solution and 8 mg (0.126 mmol) of sodium chloride. The mixture was then filtered with 0.1 mg of sodium chloride. The mixture was then dissolved in 1 ml of sodium chloride solution at a pH of 1 to 100 mm. The solution was then mixed with sodium nitrate and then dissolved in a solution of sodium nitrate and sodium nitrate at a pH of 1 to 72, and then dissolved in a solution of sodium nitrate and sodium nitrate at a pH of 1 to 72.

The test chemical is then applied to the test chemical.

The test chemical is used to determine the concentration of the test substance in the test medium.

Example 21 It consists predominantly of hydrocarbons having carbon numbers predominantly in the range of C1 through C5 and boiling in the range of approximately -15oC to -15oC (-70oF to -15oF).]

The title compound was produced by analogy to the synthesis in example 18 by coupling intermediate 30 with (2R) -2-amino-1- ((1,2-oxazinan-2-yl) -3-phenylpropan-1-one trifluoroacetic acid salt (intermediate 33) and subsequent dilution with piperidine.

The test chemical is then applied to the test chemical.

The test chemical is then applied to the test chemical and the test chemical is then removed.

The mean values for the 1H-NMR (500 MHz, DMSO-d6) are: δ = 8.8 (m, 2H), 8.65 (m, 1H), 8.32 and 8.1 (2d, 1H), 7.3-7.1 (m, 5H), 5.05 and 4.95 (2m, 1H), 4.65 (m, 1H), 4.62 and 4.56 (2t, 1H), 4.1 to 3.75 (m, 5H), 3.7-3.45 (m, 4H), 3.28, 3.22, 3.18, 3.17, 3.04 and 2.99 (6s, 9H), 2.9 and 2.75 (2m, 2H), 2.46 (m, 3H), 2.45 to 2.2 (m, 3H), 2.1-1.5 (m, 12H), 1.0 to 0.8 (m, 18H), 0.8 and 0.75 (2d, 3H) [H2O peak signals are further hidden].

Example 22 N-methyl-L-valyl-N-[(3R,4S,5S) -3-methoxy-1-{(2S)-2-[(1R,2R) -1-methoxy-2-methyl-3-{([(2S) -2-methyl-1- (1,2-oxazinan-2-yl) -1-oxo-3-phenylpropan-2-yl]amino}-3-oxopropyl]pyrrolidin-1-yl}-5-methyl-1-oxoheptan-4-yl]-N-methyl-L-valinamide Trifluoroacetic acid salt

The title compound was prepared by analogy to the synthesis in example 18 by coupling intermediate 30 with (2S) -2-amino-2-methyl-1- ((1,2-oxazinan-2-yl) -3-phenylpropan-1-one trifluoroacetic acid salt (intermediate 34) and subsequent defusion with piperidine.

The test chemical is used to determine the concentration of the test substance in the test medium.

Example 23 The substance is to be classified in the immediate vicinity of the product and the product must be classified in the immediate vicinity of the product.

50 mg (0.054 mmol) N-Methyl-L-valyl-N-[(3R,4R,5S) -3-methoxy-1-N-trifluoroacetic acid salt (Example 17) were reconstituted in 8 ml dioxy- (2S) -2- (1:1) and then reconstituted in water with 70 ml (0.108 mmol) of a 15% solution of 4-oxobutanic acid. A further reconstitution was carried out at approximately 1 μl (0.108 mmol) of 4-oxobutanic acid. This mixture was reconstituted at approximately 100 μl (0.109 mmol) of N-methoxy-hydrobutanic acid. These residues were reconstituted at approximately 300 mg (0.075 mmol) of N-methoxy-hydrobutanic acid. These residues were then reconstituted at approximately 300 mg (0.051 mmol) of N-methoxy-hydrobutanic acid. These residues were reconstituted at approximately 300 mg (0.070 mmol) of N-methoxy-hydrobutanic acid. These residues were then reconstituted at approximately 300 mg (0.053 mmol) of N-methoxy-hydrobutanic acid. These residues were then reconstituted at approximately 300 mg (0.0108 mmol) of N-methoxy-hydrobutanic acid. These residues were reconstituted at approximately 300 mg (0.0109 mmol) and then reconstituted at approximately 300 mg (0.0103 mmol) of N-hydrobutanic acid. These residues were reconstituted at approximately 300 mg (0.0103 mmol) and then reconstituted at approximately 300 mg (0.053 mmol) of N-hydrobutanic acid. These residues were reconstituted at approximately 300 mg (0.010 mmol) and then reconstituted at approximately 300 mg (0.0103 mmol) N-hydrobutanic acid. These residues were reconstituted at approximately 300 mg (0.0103 mg (0.010 mmol) and then reconstituted at approximately 300 mg N-hydrobutanic acid. These residues were reconstituted at approximately 300 mg N-hydrobutanic acid. These residues were reconstituted at approximately 300 mg (0.0103 and then recon

The test chemical is then applied to the test chemical.

The test chemical is then applied to the test chemical and the test chemical is then removed.

The 1H-NMR (500 MHz, DMSO-d6): δ = 8.95 and 8.8 (2m, 1H), 8.88 and 8.65 (2s, 1H), 7.4-7.1 (m, 5H), 5.0, 4.78, 4.65 and 4.55 (4m, 2H), 4.1 to 3.7 (m, 5H), 3.32, 3.29, 3.20, 3.12, 3.1 and 3.0 (6s, 9H), 2.75 (m, 2H), 2.63 (t, 1H), 2.4-2.2 (m, 4H), 2.1-1.2 (m, 12H), 1.2-0.8 (m, 16H), 0.75 (m, 3H) [further signals are hidden under H2O and DMSO peaks].

Example 24 The substance is to be classified in the immediate vicinity of the product and the product must be classified in the immediate vicinity of the product.

The title compound was prepared by analogy to the synthesis in example 23 by translocation of 50 mg N-Methyl-L-valyl-N-[(3R,4S,5S) -3-methoxy-1-{(2S)-2-[(1R,2R) -1-methoxy-2-methyl-3-{(2S)-1-(1,2-oxazinan-2-yl) -1-oxo-3-phenylpropan-2-yl]amino}-3-oxopropyl]pyrrolid-1-inyl}-5-methyl-1-oxoheptan-4-yl]-N-methyl-L-valinamide trifluoroacetic acid salt (Example 1) with 4-oxobutanesoic acid.

The following information is provided for the purpose of the calculation of the yield:

The test chemical is then applied to the test chemical.

The test chemical is used to determine the concentration of the test substance in the test medium.

Example 25 The following substances are to be classified in the same category as the active substance:

The title compound was prepared by analogy to the synthesis in example 23 by conversion of 15 mg N-Methyl-L-valyl-N-[(3R,4S,5S) -3-methoxy-1-{(2S)-2-[(1R,2R) -1-methoxy-2-methyl-3-{(1S,2R)-1- (1,2-oxazinane-2-ylcarbonyl) -2-phenylcyclopropyl]amino}-3-oxopropyl]pyrrolidin-1-yl}-5-methyl-1-oxoheptan-4-yl]-N-methyl-L-valinamide trifluoroacetic acid salt (Example 17) with 4-melubizoic acid.

The yield is 7.5 mg (48% d.j.)

The test chemical is then applied to the test chemical.

The test chemical is used to determine the concentration of the test substance in the test medium.

Example 26 The substance is to be classified in the immediate vicinity of the product and the product must be classified in the immediate vicinity of the product.

10 mg (0.011 mmol) N-Methyl-L-valyl-N-[(3R,4S,5S) -3-methoxy-1-N-dioxy-acid solution (Example 17) were reconstituted in 2 ml of dioxin/water (1:1) and then reconstituted with 2.8 mg (0.022 mmol) 6-methoxy-acid solution. The solution was then further reduced to 0.75 mg (0.075 mmol) by 0.75 mg (0.75 mmol) of N-dioxy-acid solution. These were then reconstituted at a pH of 6.4 °C and then further diluted at a pH of 0.75 mg (0.012 mmol) by 0.75 mg (0.75 mmol) of N-dioxy-acid solution. These were then reconstituted at a pH of 0.12 °C. The solution was then added to 0.75 mg (0.012 mmol) of N-dioxy-acid solution. These were then added to a 100 mg (0.012 mmol) of N-dioxy-acid solution and then reconstituted at a pH of 0.12 °C. These were added to a solution of 0.12 mg (0.012 mmol) of N-dioxy-acid solution and then reconstituted at a pH of 0.12 °C. These were added to a solution of 0.12 mg (0.012 mmol) N-dioxy-acid solution and then reconstituted at a pH of 0.12 °C. These were then reconstituted at a further 0.8 mg (0.012 mmol) N-dioxy-acid solution and then reconstituted at a pH of 0.12 °C. These were added to a solution of 0.12 mmol-dioxy-acidine at a solution of 0.8 mg (0.0 mg (0.012 mmol) N-acidine. These were then reconstituted at a sodium at a pH of 0.0 mg (0.012 mmol.

The test chemical is then applied to the test chemical.

The test chemical is used to determine the concentration of the test substance in the test medium.

Example 27 N- ((2-aminoethyl) -N-methyl-L-valyl-N- ((3-R,4S,5S) -3-methoxy-1-{(2S)-2-[(1R,2R) -1-methoxy-2-methyl-3-{(2-O-oxy-propane-2-yl) -1-amino-oxopropyl) -3-pyrrolidin-1-yl) -5-methyl-1-oxoheptane-4-yl) -N-methyl-L-valinamide (trifluorous bisulfuric acid) - salt of the product

The title compound was produced by analogy to the synthesis in example 2 by translocation of 68 mg N-Methyl-L-valyl-N-[(3R,4S,5S) -3-methoxy-1-{(2S)-2-[(1R,2R) -1-methoxy-2-methyl-3-{(2S)-1-(1,2-oxazinan-2-yl) -1-oxo-3-phenylpropan-2-yl]anmino}-3-oxopropyl]pyrrolid-1-inyl}-5-methyl-1-oxo-heptan-4-yl]-N-methyl-L-valinamide trifluoric acid salt (Example 1) with tert-butyl-butyl-2-oxo-ethyl) carbamate and subsequent cleavage of the Boc protection group with Trifluoric acid.

yield: 49 mg (62% d. th. over two steps)

The test chemical is then applied to the test chemical.

The test chemical is then applied to the test chemical and the test chemical is then removed.

The mean values for the 1H-NMR (600 MHz, DMSO-d6): δ = 8.25 (m, 1H), 8.45 and 8.15 (2d, 1H), 7.65-7.55 (m, 3H), 7.23-7.1 (m, 5H), 5.12 and 4.95 (2m, 1H), 4.72 and 4.62 (2m, 1H), 4.6 and 4.52 (2t, 1H), 4.2-3.8 (m, 4H), 3.7 (m, 4H), 3.23, 3.20, 3.19, 3.18, 3.03 and 2.98 (6s, 9H), 3.0-2.7 (m, 6H), 2.4-1.2 (m, 15H), 1.05, 1.0, 0.88 and 0.82 (4d, 6H), 0.92 (m, 6H), 0.73 (m, 6H) [further signals are hidden under H2O peak].

Example 28 The substance is to be classified in the immediate vicinity of the product and the product must be classified in the immediate vicinity of the product.

The title compound was synthesized by analogy to example 27 by translocation of 25 mg (0.027 mmol) N-Methyl-L-valyl-N-[(3R,4S,5S) -3-methoxy-1-{(2S)-2-[(1R,2R) -1-methoxy-2-methyl-3-{(1S,2R) -1-1,2-oxazinan-2-ylcarbonyl)-2-phenylcyclopropyl]amino}-3-oxopropyl]pyrrolidin-1-yl}-5-methyl-1-oxoheptan-4-yl]-N-methyl-L-valinamide trifluoroacetic acid salt (Example 17) with benzyl- ((3-oxopropyl) hydrogen carbamat and subsequent electrolytic cleavage of the protective group (palladium on 10% carbon as a catalyst) as an ethanol solvent.

yield: 11 mg (41% of the total over two steps)

The test chemical is then applied to the test chemical.

The test chemical is used to determine the concentration of the test substance in the test medium.

Example 29 The substance is to be classified in the immediate vicinity of the product and the product must be classified in the immediate vicinity of the product.

The title compound was prepared by analogy to the synthesis in example 23 by conversion of 9.5 mg N-Methyl-L-valyl-N-[(3R,4S,5S) -3-methoxy-1-{(2S)-2-[(1R,2R) -1-methoxy-2-methyl-3-{(2S)-1-(1,2-oxazinan-2-yl) -1-oxo-3-phenylpropan-2-yl]amino}-3-oxopropyl]pyrrolidin-1-yl}-5-methyl-1-oxoheptan-4-yl]-N-methyl-L-valinamide trifluoric acid salt (Example 1) with methyl-4-oxo-butanoate.

The yield is 4 mg (43% d.j.)

The test chemical is then applied to the test chemical.

The test chemical is used to determine the concentration of the test substance in the test medium.

Example 30 The substance is to be classified in the immediate vicinity of the product and the product must be classified in the immediate vicinity of the product.

The title compound was produced by analogy to the synthesis in Example 28 by translocation of 20 mg (16 μmol) N-Methyl-L-valyl-N-[(3R,4S,5S) -3-methoxy-1-{(2S)-2-[(1R,2R) -1-methoxy-2-methyl-3-{(2S)-1-(1,2-oxazinan-2-yl) -1-oxo-3-phenylpropan-2-yl]amino}-3-oxopropyl]pyrrolidin-1-yl}-5-methyl-1-oxoheptan-4-yl]-N-methyl-L-valyl trifluoric acid (Example 1) with benzyl6-oxohexyl) carbamate and hydrogen following the hydrolythic cleavage of the protective group (with 10% carbon dioxide as catalyst in methanol).

yield: 7.6 mg (55% of the total over two stages)

The test chemical is then applied to the test chemical.

The test chemical is used to determine the concentration of the test substance in the test medium.

Example 31 The substance is to be classified in the immediate vicinity of the product and the product must be classified in the immediate vicinity of the product.

The title compound was synthesised by analogy to example 28 by translocation of 200 mg (0.108 mmol) N-Methyl-L-valyl-N-[(3R,4S,5S) -3-methoxy-1-{(2S)-2-[(1R,2R) -1-methoxy-2-methyl-3-{(1S,2R) -1-1,2-oxazinan-2-ylcarbonyl)-2-phenylcyclopropyl]amino}-3-oxopropyl]pyrrolidin-1-yl}-5-methyl-1-oxoheptan-4-yl]-N-methyl-L-valinamide trifluoroacetic acid salt (Example 17) with benzyl- ((6-oxohexyl) carbamate and hydrogen following the electrolytic cleavage of the protective group (with 5% diethylene on coal, in methanol as a catalyst) as a solvent.

yield: 69 mg (65% d. th. over two steps)

The test chemical is then applied to the test chemical.

The test chemical is used to determine the concentration of the test substance in the test medium.

B. Evaluation of the biological efficacy

The biological activity of the compounds of the invention may be demonstrated by in vitro and in vivo studies as known to the practitioner, for example, the pharmacological and pharmacokinetic properties of the compounds of the invention may be determined by the assays described below:

B-1. Determination of antiproliferative effect on the 786-O RCC cell line:

A defined number of cells from the human renal cancer cell line 786-O were sown in a 96-well microtiter plate in a full medium (2 500 or 7 000 cells/well) and incubated overnight at 37°C / 5% CO2. After 18 h, the sowing medium was replaced by a serum-free medium or a medium with 2% FCS. Treatment started with the addition of the respective test substance in varying concentrations (10-5 M to 10-14 M). Incubation times ranged from 48 h to 96 h. Proliferation was measured by the MTT assay (ATCC, Manassas, Virginia, USA, catalog No. 30-1010K). After the incubation period, the MTT reagent was obtained for the incubation of the drug with a success rate of 4 h but was not detected by the drug's active substance. The results were obtained in two experiments with at least 570 nm, but were not identical.

The IC50 values of representative samples from this assay are given in Table 1: Other

1	4
4	14
6	0.5
8	2.4
9	3
11	1.2
12	0.2
13	4
16	8
17	4
23	11
24	5
26	18

In comparison, monomethylauristatin F (MMAF) has an IC50 of 260 nM in this test.

B-2. Determination of antiproliferative effect on the HT29wt cell line:

A defined cell count of the human colon cancer cell line HT29wt (wild type) was sown in a 96-well microtiter plate in a full medium (10% FCS-RPMI) (2 500 cells/well) and incubated overnight at 37°C / 5% CO2. After 18 h, the sowing medium was replaced with fresh medium containing 10% FCS. Treatment was started with the addition of the respective test substance. Of the substances to be tested, dose curves were determined in a concentration range of 10-5 M to 10-10-14 M (1 dilution series). Incubation times were selected from 48 h to 96 h. Detection of proliferation was achieved by the MTT assay (MATCC, Virginia, USA) (the test was successfully performed with at least two experiments, but the results were not identical to the test results obtained with the MTT assay). The test was conducted with at least 410 nm of the treated substance, but the results were obtained in two experiments using the MTT assay (NATCC, Virginia, USA).

The IC50 values of representative samples from this assay are given in Table 2 below:

1	0.1
2	0.5
4	0.2
9	1.4
17	0.5
18	1.2
23	0.7
24	1.5
29	0.1

In comparison, monomethylauristatin F (MMAF) has an IC50 of 10 nM in this test.

B3. Determination of the effect on tubulin polymerization:

The regulated up and down of microtubules allows for the precise division of chromosomes into daughter cells and is a continuously dynamic process. A disruption of this dynamic leads to faulty cell division and ultimately cell death. The increased cell division of cancer cells also makes them particularly susceptible to spindle fiber compounds, which are a fixed component of chemotherapy. Spindle fiber compounds such as paclitaxel or ectopulite lead to a greatly increased polymerization rate, while the polymerization rate of a polymerization compound such as vinca-E or ectyloquinone is greatly reduced.

Err1:Expecting ',' delimiter: line 1 column 96 (char 95)

For this assay, the test substances dissolved in DMSO were diluted in water from their initial concentration of 10 mM to 1 μM. In addition to the buffer control, the assay control included the polymerization enhancer paclitaxel and the polymerization inhibitor vinblastine. 96-well perforated plates with half a floor area were used for the measurement. The kinetics of the tubulin polymerization was monitored for 1 h at 37 °C in a longitudinal fluorescent meter. The excitation wavelengths were 355 nm, the emission was monitored at 460 nm. For the range of linear rise within the first 10 minutes, the fluorescence rate per minute (F/Δmin) was calculated, which represents the polymerization rate. The potential reduction rate of the microtubular fluorescent test substance was quantified.

B-4. Determination of plasma stability in vitro: Method A:

20 μl of this solution was removed and administered at 1 ml of rat or human plasma (male Wistar rat plasma with Li-heparin, Fa. Harlan & Winkelmann or human white-cell depleted fresh plasma from whole blood) at 37°C. From this heavily shaken plasma solution, 100 μl of aliquot was taken immediately after addition of the sample (reference value) and then at 5, 10, 30, 60, 120, 180 and 240 minutes and, if necessary, 24 hours after each 100 μl aliquot and administered in 300 μl of Acetylpyridine.

HPLC method in rat plasma:

The test chemical is used to determine the concentration of the active substance in the test chemical and to determine the concentration of the active substance in the test chemical.

HPLC method for human plasma:

The test chemical is used to determine the concentration of the active substance in the test chemical and to determine the concentration of the active substance in the test chemical.

Method B:

The test substance was incubated in rat plasma at 37°C or human plasma for 5 h under light stirring, with a 100 μl Aliquota taken at different times (0, 2, 5, 10, 20, 30, 60, 120, 180 and 300 minutes) after addition of an internal standard (10 μl), the proteins were cut by adding 200 μl of acetonitrile and the mixture was centrifuged in an Eppendorf centrifuge for 5 minutes after adding 150 μl of ammonium acetate buffer at pH 3 to 150 μl of the residue, the unchanged test substance was analysed by LC/MS.

The mean half-lives (t1/2) of representative samples in rat plasma obtained from these data are given in Table 3: Other

4	3	B
16	0.75	A
17	>24	A
18	>24	A
22	2	A
23	>24	A

In comparison, the methyl ester of monomethylauristatine F (MMAF-OMe) in rat plasma has a t1/ 2 value (method A) of < 1 min.

In human plasma, as an example of the compounds of the invention, no degradation was observed after 24 h in Examples 1, 17 and 21, while the methyl ester of monomethylauristatine F (MMAF-OMe) was degraded by about 20% during this period.

The test chemical is used to determine the concentration of the test chemical in the test medium.

The cell permeability of a substance can be studied by in vitro testing in a flux assay using caco-2 cells [M.D. Troutman and D.R. Thakker, Pharm. Res. 20 (8), 1210-1224 (2003) ], where the cells were cultured on 24-hole filter plates for 15-16 days, and permeation was determined by applying the respective test substance in a HEPES buffer either apical (A) or basal (B) to the cells and incubating for 2 h. After 0 h and after 2 h, samples were taken from the cis and trans probes. The samples were transported by HPLC (Agilent 1200, Böbkom, Germany) using a paper spray. The system was published on a 4-phase basis on a 4-phase basis. The test was calculated on a tri-phase basis. The test was performed on a turbo-metallic ion (A.T.A.T.A.T.A.T.A.T.A.T.A.) with a molecular weight of < 4000 μm2 (Applied to the test chemical) and was classified as a 4-phase ion (Applied to the test chemical) with a pH of > 104.5 (Applied to the test chemical) and a pH of < 104.5 (Applied to the test chemical) was classified as a 4-phase ion (Applied to the test chemical) and a 4-phase ion (Applied to the test chemical) was classified as a 4-phase ion (Applied to the test chemical) and a 4-phase ion (Applied to the test chemical) at a 4-phase).

B-6. Determination of the substrate properties of P-glycoprotein (P-gp):

Many tumour cells express transport proteins for active substances, often with the development of resistance to cytotoxic agents, and substances that are not substrates of such transport proteins, such as P-glycoprotein (P-gp) or BCRP, may therefore have an enhanced activity profile.

The substrate properties of a substance for P-gp (ABCB1) were determined by a flux assay using P-gp-over-expressing LLC-PK1 cells (L-MDR1 cells) in either a HES-buffered API (A) or basal (B) and applied to the papules (A.H. Schinkel et al., J. Clin. Invest. 96, 1698-1705 (1995)). To this end, the LLC-PK1 or L-MDR1 cells were cultured on 96-hole filter plates for 3-4 days. To determine permeation, the respective test substance was administered alone or in the presence of an inhibitor (such as ivermectin or verapamil) in a HES-buffered API (A) or basal (B) and applied to the papules using a 2 HP AP (A) or a 2 HP AP (B) and published in the journal Papyrus. The results were calculated using a 2 HP and 2 HP 0 (B) phase transducer system. The results were obtained using a Cis-SPS system, which was classified as a 3-substrate test chemical (A) and was used on a PLC-B (A) and a PLC-B (B) substrate (A) (A) (A) (A) (A) (A) (A) (A) (A) (A) (A) (A) (A) (A) (A) (A) (A) (A) (A) (A) (A) (A) (A) (A) (A) (A) (A) (A) (A) (A) (A) (A) (A) (A) (A) (A) (A) (A) (A) (A) (A) (A) (A) (A) (A) (A) (A) (A) (A) (A) (A) (A) (A) (A) (A) (A) (A) (A) (A) (A) (A) (A) (A) (A) (A) (A) (A) (A) (A) (A) (A) (A) (A)

Further criteria for assessing P-gp substrate properties may include comparison of efflux ratios in L-MDR1 and LLC PK1 cells or efflux ratios in the presence or absence of an inhibitor.

C. Examples of pharmaceutical formulations

The compounds of the invention may be converted into pharmaceutical preparations as follows:

The tablets: The composition:

100 mg of the compound of the invention, 50 mg of lactose (monohydrate), 50 mg of cornstarch (native), 10 mg of polyvinylpyrrolidone (PVP 25) (BASF, Ludwigshafen, Germany) and 2 mg of magnesium stearate.

The tablet weighs 212 mg, has a diameter of 8 mm and a bulb radius of 12 mm.

Manufacture:

The mixture of the compound of the invention, lactose and starch is granulated with a 5% solution (m/m) of PVP in water. After drying, the granulate is mixed with the magnesium stearate for 5 minutes. This mixture is pressed with a standard tablet press (tablets format see above).

Suspension for oral use: The composition:

1000 mg of the compound of the invention, 1000 mg of ethanol (96%), 400 mg of Rhodigel® (Xanthan gum from FMC, Pennsylvania, USA) and 99 g of water.

A single dose of 100 mg of the compound of the invention corresponds to 10 ml of oral suspension.

Manufacture:

The rodigel is suspended in ethanol, the compound of the invention is added to the suspension, water is added during stirring and the rodigel is stirred for about 6 h until the fermentation is completed.

Solution for oral application: The composition:

500 mg of the compound of the invention, 2.5 g of polysorbate and 97 g of polyethylene glycol 400. A single dose of 100 mg of the compound of the invention corresponds to 20 g of oral solution.

Manufacture:

The compound of the invention is suspended in the mixture of polyethylene glycol and polysorbate under stirring and the stirring process is continued until the compound of the invention is completely dissolved.

Injection of the product:

The compound of the invention is dissolved in a physiologically compatible solvent (e.g. isotonic saline, glucose solution 5% and/or PEG 400 solution 30%) at a concentration below saturation solubility, sterilized and filled in sterile, pyrogen-free containers for injections.

Claims (14)

1. Compound of the formula (I) in which

   R¹ represents hydrogen, (C₁-C₆ alkyl) or a group of the formula Q¹-L¹-* or Q²-L²- *, wherein

   * indicates the point of attachment with the nitrogen atom,

   Q¹ denotes hydroxycarbonyl, (C₁-C₄) alkoxycarbonyl or benzyloxycarbonyl,

   L¹ denotes straight-chain (C₁-C₁₂) alkanediyl which may be substituted up to four times with methyl and in which (a) two carbon atoms in 1,2 relation, 1,3 relation or 1,4 relation to one another including the carbon atoms optionally lying between them may be bridged to form a (C₃-C₆) cycloalkyl ring or a phenyl ring or (b) up to three CH₂ groups not adjacent to one another may be exchanged for -O-,

   Q² denotes hydroxy, amino or mono-(C₁-C₄) alkylamino,

   and

   L² denotes straight-chain (C₂-C₁₂) alkanediyl which may be substituted up to four times with methyl and in which (a) two carbon atoms in 1,2 relation, 1,3 relation or 1,4 relation to one another including the carbon atoms optionally lying between them may be bridged to form a (C₃-C₆) cycloalkyl ring or a phenyl ring or (b) up to three CH₂ groups not adjacent to one another may be exchanged for -O-,

   R² represents methyl or hydroxy,

   R³ represents hydrogen or methyl,

   R⁴ represents isopropyl, isobutyl, sec.-butyl, tert.-butyl, phenyl, benzyl, 4-hydroxybenzyl, 4-hydroxy-3-nitrobenzyl, 1-phenylethyl, diphenylmethyl, 1H-imidazol-4-ylmethyl or 1H-indol-3-ylmethyl,

   or R³ and R⁴ together with the carbon atom, to which they are both bound, form a 2-phenylcyclopropane-1,1-diyl group of the formula wherein

   # indicates the points of attachment with the remaining moieties of the molecule,

   and the ring A with the N-O group present therein represents a monocyclic or bicyclic, optionally substituted heterocycle of the formula wherein

   ** indicates the point of attachment with the carbonyl group,

   R⁵ and R⁶ denote in each case hydrogen, hydroxy, (C₁-C₄) alkoxy or benzyloxy, and R⁷ denotes hydrogen, fluorine, chlorine, cyano, methyl or trifluoromethyl, and its salts, solvates and solvates of the salts.
2. Compound of the formula (I) according to claim 1, in which

   R¹ represents hydrogen, (C₁-C₄ alkyl) or a group of the formula Q¹-L¹-* or Q²-L²- *, wherein

   * indicates the point of attachment with the nitrogen atom,

   Q¹ denotes hydroxycarbonyl, (C₁-C₄) alkoxycarbonyl,

   L¹ denotes straight-chain (C₁-C₁₂) alkanediyl in which (a) two carbon atoms in 1,3 relation or 1,4 relation to one another including the one or the two carbon atoms lying between them may be bridged to form a phenyl ring or (b) up to three CH₂ groups not adjacent to one another may be exchanged for -O-,

   Q² d enotes hydroxy, amino or methylamino, and

   L² denotes straight-chain (C₂-C₁₂) alkanediyl which may be substituted once or twice with methyl and in which up to three CH₂ groups not adjacent to one another may be exchanged for -O-,

   R² represents methyl or hydroxy,

   R³ represents hydrogen,

   R⁴ represents benzyl, 4-hydroxybenzyl, 1-phenylethyl or 1H-indol-3-ylmethyl,

   or R³ and R⁴ together with the carbon atom, to which they are both bound, form a 2-phenylcyclopropane-1,1-diyl group of the formula wherein

   # indicates the points of attachment with the remaining moieties of the molecule,

   and the ring A with the N-O group present therein represents a monocyclic or bicyclic, optionally substituted heterocycle of the formula wherein

   ** indicates the point of attachment with the carbonyl group,

   and R⁵ denotes hydrogen, hydroxy or benzyloxy, and its salts, solvates and solvates of the salts.
3. Compound of the formula (I) according to claim 1 or 2, in which

   R¹ represents hydrogen, methyl or a group of the formula Q¹-L¹-* or Q²-L²- *, wherein

   * indicates the point of attachment with the nitrogen atom,

   Q¹ denotes hydroxycarbonyl, methoxycarbonyl or ethoxycarbonyl,

   L¹ denotes straight-chain (C₁-C₆) alkanediyl, in which two carbon atoms in 1,4 relation to one another including the two carbon atoms lying between them may be bridged to form a phenyl ring

   Q² denotes hydroxy or amino, and

   L² denotes straight-chain (C₂-C₆) alkanediyl,

   R² represents methyl,

   R³ represents hydrogen,

   R⁴ represents benzyl, 1-phenylethyl or 1H-indol-3-ylmethyl,

   or R³ and R⁴ together with the carbon atom, to which they are both bound, form a (1S,2R)-2-phenylcyclopropane-1,1-diyl group of the formula wherein

   #1 indicates the point of attachment with the adjoining nitrogen atom,

   and

   #2 indicate the point of attachment with the carbonyl group,

   and the ring A with the N-O group present therein represents a monocyclic or bicyclic, optionally substituted heterocycle of the formula wherein

   ** indicates the point of attachment with the carbonyl group,

   and

   R⁵ denotes hydrogen, hydroxy or benzyloxy,

   and its salts, solvates and solvates of the salts.
4. Compound according to claim 1, 2 or 3 with the formula (1-A) in which R¹, R², R³, R⁴ and the ring A have the meanings defined in claim 1,2 or 3 and the carbon atom carrying the radicals R³ and R⁴ has the S configuration shown, and its salts, solvates and solvates of the salts.
5. Compound of claim 1, wherein said compound is selected from the group consisting of: and
6. Method for producing a compound of the formula (I), as defined in claims 1 to 4, characterised in that a compound of the formula (II) in which R² has the meaning indicated in claims 1 to 4 and PG represents an amino protective group, such as for example (9H-fluoren-9-ylmethoxy)carbonyl, tert.-butoxycarbonyl or benzyloxycarbonyl, is coupled in an inert solvent with activation of the carboxyl function in (II) either

   [A] initially with the compound of the formula (III) or a salt thereof to form a compound of the formulae (IV) in which R² and PG have the meanings indicated above, and then the latter is coupled in an inert solvent with activation of the carboxyl function with a compound of the formula (V) in which R³, R⁴ and the ring A have the meanings indicated in claims 1 to 4, or a salt of this compound to form a compound of the formula (VI) in which R², R³, R⁴, the ring A and PG have the meanings indicated above, or

   [B] is coupled with a compound of the formula (VII) in which R³, R⁴ and the ring A have the meanings indicated in claims 1 to 4, or a salt of this compound likewise to form the compound of the formula (VI) in which R², R³, R⁴, the ring A and PG have the meanings indicated above, and the particular resulting compound of the formula (VI) is then de-protected by conventional methods of peptide chemistry to form a compound of the invention of the formula (1-B) in which R², R³, R⁴ and the ring A have the meanings indicated above, and the latter is then converted, if required, either (i) by base-induced alkylation with a compound of the formula (VIII)         R^1A-X     (VIII), in which

   R^1A has the meaning of R¹ indicated in claims 1 to 4, but does not represent hydrogen,

   and

   X represents a volatile group, such as for example chloride, bromide, iodide, mesylate, triflate or tosylate,

   to a compound of the invention of the formula (I-C) in which R^1A, R², R³, R⁴ and the ring A have the meanings indicated above, or (ii) is converted by reacting with a compound of the formula (IX) in which

   R^1B has the meaning of R^1A indicated above, but shortened in the alkyl chain length by one CH₂ unit,

   in the presence of a suitable reducing agent to a compound of the invention of the formula (I-D)

   in which R^1B, R², R³, R⁴ and the ring A have the meanings indicated above, and the compounds thus obtained of the formulae (1-B), (1-C) or (1-D) are separated optionally into their enantiomers and/or diastereomers and/or reacted with the corresponding (i) solvents and/or (ii) bases or acids to form their solvates, salts and/or solvates of the salts.
7. Compound as defined in one of claims 1 to 5 for use in a method for treating and/or preventing diseases.
8. Compound as defined in one of claims 1 to 5 for use in a method for treating and/or preventing carcinosis and oncosis.
9. Use of a compound as defined in one of claims 1 to 5 for producing a medicament for treating and/or preventing carcinosis and oncosis.
10. Medicament containing a compound as defined in one of claims 1 to 5 in combination with one or more inert, non-toxic, pharmaceutically suitable adjuvants.
11. Medicament containing a compound as defined in one of claims 1 to 5 in combination with one or more further active ingredients.
12. Medicament according to claim 10 or 11 for use in a method for treating and/or preventing carcinosis and oncosis.
13. Antiproliferative-acting conjugate in which the compound according to one or more of claims 1-5 is linked to a protein.
14. Antiproliferative-acting conjugate according to claim 13, wherein the protein is an antibody.