DE102009051799A1 - Bifunctional prodrugs and drugs - Google Patents

Abstract

The present invention is directed to new compounds, more particularly to novel bifunctional prodrugs and drugs. In another aspect, the present invention is directed to pharmaceutical compositions containing these prodrugs and drugs and their use as cytostatic agents in tumor therapy. The compounds according to the invention are based on CC-1065 analogues.

Description

The present invention is directed to new compounds, more particularly to novel bifunctional prodrugs and drugs. In another aspect, the present invention is directed to pharmaceutical compositions containing these prodrugs and drugs and their use as cytostatic agents in tumor therapy. The compounds according to the invention are based on CC-1065 analogues.

State of the art

The diverse manifestations of cancer require individual therapy concepts. In response to the complexity of a tumor disease, most of today's clinically applied treatments are combinations of different therapeutic approaches. On the one hand, in well-accessible and well-defined tumors, surgical removal of degenerated tissue is used as the method of choice. However, if the tumor is more difficult to access or vital to structures, radiotherapy is the method of choice. In a more advanced stage, in which metastases have already formed or at least risk metastasizing, chemotherapy is usually performed immediately. Furthermore, the methods of hormone therapy, immunotherapy and therapy with angiogenesis inhibitors and kinase inhibitors are used in tumor therapy.

In the case of metastasis already proven or suspected, as well as in the case of systemic tumors, chemotherapy is currently the most important treatment, although it is often associated with severe side effects such as blood disorder, immunodeficiency, mucositis, fever, nausea, vomiting, etc. Typically, the chemotherapeutic agents are distributed throughout the body through the bloodstream so that they can reach all cells. Chemotherapeutic agents have cytostatic effects on human cells, d. they prevent cell proliferation, or they are cytotoxic, i. that is, they cause the cells to die.

Since most cytostatics show their mechanism of action in proliferating cells and tumor cells are among the rapidly proliferating cell types, they are used as chemotherapeutic agents. However, the non-tumor cells of the treated person are also affected, especially those of the bone marrow, the hair roots or mucosal cells.

The currently known chemotherapeutic cytotoxic agents are classified according to their mechanisms of action in the classes of alkylating agents, antimetabolites, mitotic inhibitors, topoisomerase inhibitors and cytostatic antibiotics.

The alkylating agents represent a numerically significant, structurally very diverse class of extremely reactive substances. After an optionally preceding activation of the drug to the carbocation, the active ingredient reacts as electrophilic, in particular with nucleic acids to form covalent bonds. As a result, cross-linking of the DNA, abnormal base pairing or strand breaks occur that prevent replication and eventually lead to cell death. Typical examples of alkylating agents are cyclophosphamide but also cisplatin. In addition to a group of particularly effective alkylating agents include the natural antibiotic CC-1065, duocarmycins, yatakemycin and derivatives and analogues of this natural product class.

Due to the need for chemotherapeutic treatments, the severe side effects of most clinically used drugs, and the emergence of resistance to many known chemotherapeutic agents, continuous development in the field of chemotherapeutics is needed.

Today, chemotherapy for malignant disease is associated with severe side effects, as differentiation between healthy and malignant tissue occurs only through the increased proliferation rate of cancer cells. Therefore, new concepts have been developed which exploit genotypic and phenotypic properties of tumor cells and enable a targeted activation of reversible detoxified prodrug directly at the site of action. Such a targeted activation can z. B. by a changed ph value, z. As a pH reduction, done. Another possibility is the so-called ADEPT concept (Antibody-Directed Enzyme Prodrug Therapy). This antibody-enzyme conjugates are used, which causes a direct conversion to the tumor of the non-toxic prodrug in the drug and achieve high selectivity. This binary therapy approach consists of two steps. First, a certain amount of antibody-enzyme conjugate is applied, which is then distributed through the bloodstream throughout the organism. The conjugate binds to specific antigens on tumor cell surfaces or is degraded by the body or excreted. If the unbound antibody-enzyme conjugate can no longer be detected, the application of the prodrug takes place in the second step. The most non-toxic prodrug is also distributed throughout the organism and specifically targeted in the tumor tissue due to the enzyme present only on the tumor surface in the form of the antibody-enzyme conjugate. The released drug then unfolds its toxic effect upon penetration through the cell membrane, while the enzyme remains active on the outside of the tumor cell and can activate further prodrug molecules. Cleavage of the prodrugs by the body's own enzyme systems should as far as possible not take place in the context of this approach, since otherwise the activity of the therapy would be reduced or eliminated. Disadvantages of hitherto known prodrugs, however, are too low a cytotoxicity difference between the prodrug and the drug generated therefrom as well as a too low cytotoxicity of the drug itself.

As a guideline in the development of compounds for the ADEPT concept, the QIC ₅₀ value (QIC ₅₀ = IC ₅₀ (prodrug) / IC ₅₀ (prodrug in the presence of the enzyme)) should be greater than 1000 and the cytotoxicity of the basis Drugs should have an IC ₅₀ value (toxin concentration at which cell growth is suppressed by 50%) less than 10 nM.

A further approach in the context of a targeted treatment of malignant tumors is the prodrug monotherapy. This is based on the presence of overexpressed in tumors enzymes that are able to cleave a corresponding prodrug with release of the corresponding drug. One possible enzyme is z. As the β-D-glucuronidase, which could be detected in increased concentrations in necrotic areas of tumor tissue. Alternatively, conjugates of drugs and tumor-specific ligands can be used for selective targeting in cancer therapy. After selective binding of the ligand to a receptor on the tumor surface, internalization of the conjugate and, subsequently, intracellular release occur.

Other methods in addition to the ADEPT method described above are the methods known in the art PDEPT (Polymer-Directed Enzyme Prodrug Therapy) or MDEPT (Macromolecular Directed Enzyme Prodrug Therapy) and the VDEPT (Virus-Directed Enzyme Prodrug Therapy) or GDEPT (Gene -Directed Enzyme Prodrug Therapy). In particular, the latter two are ways to reinforce the above-mentioned prodrug monotherapy.

Clinical trials have already been conducted for the ADEPT concept. It turned out that the ADEPT concept per se is suitable for selective tumor therapy, but there is still room for improvement with regard to various points in order to enable a selective and efficient therapy. One of these significant improvement points is the provision of new and effective prodrugs that have a high cytotoxicity difference between the prodrug and the corresponding drug, a high cytotoxicity of the drug and a short plasma half-life of the drug.

There is therefore still a need to provide corresponding prodrugs which have a high cytotoxicity difference from the drug and, as a prodrug itself, exhibit low cytotoxicity. The drug itself should have the highest possible cytotoxicity.

For analogues of the antibiotic CC-1065 and the duocarmycines various attempts have been made to achieve the above-mentioned goal. The CC-1065 itself has an IC ₅₀ value of 20 pmol, but has led to delayed lethal hepatotoxicity in animal studies and is therefore not suitable for clinical applications. Therefore, attempts have been made to present analogs of this compound. Thus, the DNA binding unit was changed, but also the pharmacophore itself was modified in various forms. In addition, seco and prodrug compounds have been synthesized which generally exhibit similar toxicity and selectivities as the spirocyclopropyl compounds, e.g. B. CC-1065 or duocarmycins. So z. As described by Tietze CC-1065 analogs, which reversibly by glycosidation of the detoxified anti-methyl-seco-CBI-DMAI-β-D-galactoside (+) - (1S, 10R) compound an excellent result in terms of cytotoxicity and the To achieve quotients of the cytotoxicity of the prodrug and the prodrug in the presence of the activated enzyme. QIC ₅₀ values in excess of 4500 were achieved.

Bifunctional alkylating agents are those with two reactive centers. They have the ability to cause intra- or interstrand cross-links of the DNA. With regard to the interstrand cross-linking, particularly good damage to the cells and thus cell death are achieved. These compounds have a high cytotoxicity. Bifunctional derivatives of e.g. B. Pyrrolobenzodiazidpine are described in the literature. Such bifunctional derivatives are currently in Phase 1 clinical trials. Also for analogues of CC-1050 bifunctional compounds were synthesized, Mitchell et al., J. Am. Chem. Soc. 1989, 111, 6428-6429 ; Jia, G., and Lown, W., Bioorg. Med. Chem., 2000, 8, 1607-1617 , However, there is still a need for drugs and prodrugs suitable for chemotherapeutic agents.

Description of the invention

According to the invention, new compounds are provided which represent bifunctional alkylating agents in particular for use in a selective tumor therapy. The novel compounds described herein are characterized by the fact that these new dimers are more cytotoxic than the monomeric prodrugs or drugs. Usually, the IC ₅₀ value of the compounds of the invention in the pmol range. Furthermore, a much larger QIC ₅₀ value is achieved. That is, the quotient between the cytotoxicity of the drug and the cytotoxicity of the prodrug is much greater. Thus, a better therapeutic effectiveness associated with a lower cytotoxicity of the prodrug and thus lower side effects in applications in patients can be achieved.

mit R¹ ausgewählt aus einem Halogen und OSO₂R^u, wobei R^u ausgewählt ist aus gegebenenfalls substituierten C₁-C₆ alkyl, C_1-6 Perhaloalkyl, Benzyl und Phenyl;
R² ist ausgewählt aus Wasserstoff und gegebenenfalls substituierten C_1-8 Alkyl;
R³, R^3', R⁴ und R^4' sind unabhängig voneinander ausgewählt aus Wasserstoff und gegebenenfalls substituierten C_1-8 Alkyl, wobei zwei oder mehrere von R², R³, R^3', R⁴ und R^4' gegebenenfalls miteinander verbunden sind, um ein oder mehrere gegebenenfalls substituierte Kohlenstoffzyklen oder Heterozyklen auszubilden;
X² ist ausgewählt aus O, C(R¹⁴)(R^14') und NR^14', wobei R¹⁴ ausgewählt ist aus Wasserstoff und gegebenenfalls substituierten C_1-8 Alkyl oder C_1-8 Acyl und wobei R^14' nicht vorhanden ist oder ausgewählt ist aus Wasserstoff und gegebenenfalls substituierten C_1-8 Alkyl oder C_1-8 Acyl;
R⁵, R^5', R⁶, R^6', R⁷ und R^7' sind unabhängig voneinander ausgewählt aus H, OH, SH, CF₃, CN, C(O)NH₂, C(O)H, C(O)OH, Halogen, R^k, SR^k, S(O)R^k, S(O)₂R^k, S(O)OR^k, S(O)₂OR^k, OS(O)R^k, OS(O)₂R^k, OS(O)OR^k, OS(O)₂R^k, OR^k, P(O)(OR^k)(OR^L), OP(O)(OR^k)(OR^L), SiR^kR^LR^m, C(O)R^k, C(O)OR^k, C(O)N(R^L)R^k, OC(O)R^k, OC(O)R^k, OC(O)OR^k, OC(O)N(R^k)R^L wobei R^k, R^L und R^m unabhängig voneinander ausgewählt sind aus H und gegebenenfalls substituierte C_1-4 Alkyl, C_1-4 Heteroalkyl, C_3-7 Cycloalkyl, C_3-7 Heterocycloalkyl, C_4-12 Aryl oder C_4-12 Heteroaryl Gruppen, zwei oder mehr von R^k, R^L und R^m bilden gegebenenfalls miteinander ein oder mehrere gegebenenfalls substituierte aliphatische oder aromatische Kohlenstoffzyklen oder Heterozyklen aus;
und/oder R^5' und R^6' und/oder R^6' und R^7' und/oder R^7' und R^14' sind nicht vorhanden, d. h., sie bilden eine im Ring vorhandene Doppelbildung zwischen den mit den entsprechenden substituierten Atomen aus, nämlich R^5' und R^6', und/oder R^6' und R^7', und/oder R^7' und R^14';
zwei oder mehrere von R⁵, R^5', R^6', R^7', R¹⁴, und R^14' können gegebenenfalls miteinander verbunden sein, um ein oder mehrere gegebenenfalls substituierte aliphatische oder aromatische Kohlenstoffzyklen oder Heterozyklen auszubilden;
und/oder R⁵ + R^5', und/oder R⁶ + R^6' und/oder R⁷ + R^7' sind unabhängig voneinander =O, =S oder =NR¹², wobei R¹² ausgewählt ist aus H und gegebenenfalls substituierten C_1-6 Alkyl;
X₆ ist ausgewählt aus C oder N;
R_DB ist ausgewählt aus Wasserstoff und gegebenenfalls substituierten C_1-8 Alkyl oder gegebenenfalls substituierten C_1-8 Acyl;
X₁ ist ausgewählt aus O, S, und NR¹³, wobei R¹³ ausgewählt ist aus H und gegebenenfalls substituierten C_1-8 Alkyl;
R ist ausgewählt aus Wasserstoff oder ein enzymatisch abspaltbares Substrat;
a und b sind unabhängig voneinander ausgewählt aus 0 und 1;
c ist ausgewählt aus 0, 1 und 2;
d ist ausgewählt aus 0 oder 1;
L ist eine Verbindungsgruppe zur kovalenten Verbindung von A und B; und pharmazeutisch annehmbare Salze oder pharmazeutisch annehmbare Solvate hiervon.In a first aspect, the present invention provides compounds of the general structure
ALB
wherein A and B are independently selected from the structures I or II

wherein R ^{1 is} selected from a halogen and OSO ₂ R ^u , wherein R ^{u is} selected from optionally substituted C ₁ -C ₆ alkyl, C _1-6 perhaloalkyl, benzyl and phenyl;
R ² is selected from hydrogen and optionally substituted C _1-8 alkyl;
R ³ , R ^{3 '} , R ⁴ and R ^4' are independently selected from hydrogen and optionally substituted C _1-8 alkyl, wherein two or more of R ² , R ³ , R ^{3 '} , R ⁴ and R ^{4' are} optionally linked together to form one or more optionally substituted carbon cycles or heterocycles;
X ² is selected from O, C (R ¹⁴ ) (R ^{14 '} ) and NR ^14' , wherein R ^{14 is} selected from hydrogen and optionally substituted C _1-8 alkyl or C _1-8 acyl and where R ^{14 '} is absent is or is selected from hydrogen and optionally substituted C _1-8 alkyl or C _1-8 acyl;
R ⁵ , R ^{5 '} , R ⁶ , R ^6' , R ⁷ and R ^{7 '} are independently selected from H, OH, SH, CF ₃ , CN, C (O) NH ₂ , C (O) H, C (O) OH, halogen, R ^k , SR ^k , S (O) R ^k , S (O) ₂ R ^k , S (O) OR ^k , S (O) ₂ OR ^k , OS (O) R ^k , OS (O) ₂ R ^k , OS (O) OR ^k , OS (O) ₂ R ^k , OR ^k , P (O) (OR ^k ) (OR ^L ), OP (O) (OR ^k ) (OR ^L ), SiR ^k R ^L R ^m , C (O) R ^k , C (O) OR ^k , C (O) N (R ^L ) R ^k , OC (O) R ^k , OC (O) R ^k , OC (O) OR ^k , OC (O) N (R ^k ) R ^L where R ^k , R ^L and R ^{m are} independently selected from H and optionally substituted C _1-4 alkyl, C _1-4 heteroalkyl, C _{3- 7} cycloalkyl, C _3-7 heterocycloalkyl, C _4-12 aryl or C _4-12 heteroaryl groups, two or more of R ^k , R ^L and R ^m optionally together form one or more optionally substituted aliphatic or aromatic carbon cycles or heterocycles;
and / or R ^{5 '} and R ^6' and / or R ^{6 '} and R ^7' and / or R ^{7 '} and R ^14' are absent, ie they form a double ring present in the ring between those with the corresponding substituted atoms from, namely R ^{5 '} and R ^6' , and / or R ^{6 '} and R ^7' , and / or R ^{7 '} and R ^14' ;
two or more of R ^5, R ^{5 ',} R ^6', R ^{7 ',} R ^14, and R ^14' may optionally be connected together to form one or more optionally substituted aliphatic or aromatic carbon rings or heterocycles;
and / or R ⁵ + R ^{5 '} , and / or R ⁶ + R ^6' and / or R ⁷ + R ^{7 '} are independently = O, = S or = NR ¹² , wherein R ^{12 is} selected from H and optionally substituted C _1-6 alkyl;
X ₆ is selected from C or N;
R _DB is selected from hydrogen and optionally substituted C _1-8 alkyl or optionally substituted C _1-8 acyl;
X ₁ is selected from O, S, and NR ¹³ , wherein R ^{13 is} selected from H and optionally substituted C _1-8 alkyl;
R is selected from hydrogen or an enzymatically cleavable substrate;
a and b are independently selected from 0 and 1;
c is selected from 0, 1 and 2;
d is selected from 0 or 1;
L is a linking group for the covalent linking of A and B; and pharmaceutically acceptable salts or pharmaceutically acceptable solvates thereof.

The compounds of the invention showed in vitro experiments excellent cytotoxicity with IC ₅₀ values in the pmol range, partially below the pmol range. Furthermore, the compounds showed an excellent quotient of the IC ₅₀ . The QIC ₅₀ value of the tested compounds was over 1000, especially suitable compounds showed values over 100,000.

That is, the present compounds, which are novel dimeric prodrugs and drugs of CC-1065 analogues, have a very high cytotoxicity as a drug, while the prodrugs show less cytotoxicity. Thus, these compounds should be much safer in therapeutic use. The compounds according to the invention from diners are furthermore substantially more cytotoxic than the monomeric prodrugs or drugs. The compounds are thus characterized by a high efficacy with fewer side effects in the application.

Earlier bifunctional CC-1065 and duocarmycin analogues showed only low selectivity and also the cytotoxicity values were similar to the monomeric analogs, sometimes even lower. Jia, G. and Lown, J.W., supra, examined bifunctional compounds that showed cytotoxicity values in the nanomolar range.

The compounds of the invention are characterized in that they exhibit cytotoxicity values in the pmol range and have a very large quotient of the IC ₅₀ values of the drug to the prodrug.

In a preferred embodiment, the compounds have a linker L selected from i) ZC (R _CH ) ₂ ) _n -Z ', where n is an integer from 1 to 20, Z and Z' are independently selected from C = O, OC = O, SO ₂ , NR _z , NR _z C = O, C = ONR _z wherein each R _CH and R _{z are} independently selected from hydrogen or optionally substituted alkyl or optionally substituted acyl, preferably from optionally substituted C ₁ C ₈ alkyl or optionally substituted C ₁ -C ₈ acyl, or ii) L is an oligopeptide of 1 to 10 amino acids; or pharmaceutically acceptable salts or substrates thereof.

In a preferred embodiment, the linker is one wherein Z, Z 'is a group C = O and R _{CH is} each independently hydrogen. In one embodiment, n is from 1 to 6, more preferably 3, 4 or 5. In another embodiment, each R _CH is independently hydrogen or CH ₃ , n is an integer from 1 to 20 and Z and Z 'are C = O, where X ^{6 is} nitrogen.

Furthermore, it is preferred that in the general structure I a is zero.

A further preferred embodiment is such that in the structures I and II the radicals R ⁶ , R ^{6 '} , R ⁷ , R ^7' and R ^{2 are} each independently of one another a hydrogen or a methyl group -CH ₃ -.

In a further preferred embodiment, the structures I and II are those CC-1065 analogs, as z. B. in the WO 2007/089149 and WO 01/83448 are fully incorporated herein by reference.

The term "substituted", as used herein in particular with reference to alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, acyl, refers to those groups having one or more substituents selected from the group comprising OH, = O, = S, = NR ^h , = N-OR ^h , S ^h , NH ₂ , NO ₂ , NO, N ₃ , CF ₃ , CN, OCN, SCN, NCO, NCS, C (O) NH ₂ , C (O ) H, C (O) OH, halo, R ^h , SR ^h , S (O) R ^h , S (O) OR ^h , S (O) ₂ R ^h , S (O) ₂ OR ^h , OS (O R ^h , OS (O) OR ^h , OS (O) ₂ R ^h , OS (O) ₂ OR ^h , OP (O) (OR ^h ) (OR ⁱ ), P (O) (OR ^h ) (OR ⁱ ), OR ^h , NHR ⁱ , N (R ^h ) R ⁱ , ⁺ N (R ^h ) (R ⁱ ) R ^j , Si (R ^h ) (R ⁱ ) R ^j , Si (R ^h ) (R ⁱ (R ^j ), C (O) R ^h , C (O) OR ^h , C (O) N (R ⁱ ) R ^h , OC (O) R ^h , OC (O) OR ^h , OC (O) N (R ^h ) R ⁱ , N (R ⁱ ) C (O) R ^h , N (R ⁱ ) C (O) OR ^h , N (R ⁱ ) C (O) N (R ^j ) R ^h , and the thio derivatives of these substituents, or a protonated or deprotonated form of these substituents, wherein R ^h , R ⁱ , and R ^{j are} independently selected from H and optionally substituted C _{1- 15 is} alkyl, C _1-15 heteroalkyl, C _3-15 cycloalkyl, C _3-15 heterocycloalkyl, C _4-15 aryl, or C _4-15 heteroaryl or a combination thereof, two or more of R ^h , R ⁱ , and R ^{Optionally, j} are joined together to form one or more carbon cycles or heterocycles.

The term "alkyl" as used herein refers to straight-chained or branched, saturated or unsaturated hydrocarbon substituents, examples of the alkyl groups include methyl, ethyl, propyl, butyl, pentyl, hexyl, octyl, decyl, isopropyl, sec Butyl, isobutyl, tert-butyl, isopentyl, vinyl, allyl, 1-butenyl, 2-butenyl, isobutenyl, pentenyl and the like.

The term "cycloalkyl" or "carbon cycles" as used herein refers to saturated or unsaturated, non-aromatic hydrocarbon cycles which may consist of one, two or more rings. Examples thereof include cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclopentadienyl, cyclohexyl, cyclohexonyl, etc.

The term "heteroalkyl" as used herein refers to straight-chain or branched, saturated or unsaturated hydrocarbon substituents in which at least one carbon is replaced by a heteroatom. The heteroatoms are preferably selected from S, N, O, and P.

The term "aryl" as used herein refers to aromatic substituents which may be one or more rings fused together. Examples of aryl include phenyl, naphthyl and antracenyl.

The term "heteroaryl" as used herein refers to aromatic substituents which may consist of one or more rings fused together. Here, at least one carbon atom of the aromatic R group is replaced by a heteroatom. Examples of heteroaryl groups include pyridinyl, furanyl, pyrrolyl, triazolyl, pyrazolyl, imidazolyl, thiophenyl, indolyl, benzofuranyl, benzimidazolyl, indazolyl, benzotriazolyl, benzisoxazolyl, and quinolinyl.

The term "heterocycloalkyl" or "heterocycles" as used herein refers to saturated or unsaturated, non-aromatic cyclic hydrocarbon substituents which may consist of one or more fused rings, with at least one carbon in one of the rings replaced by a heteroatom. Examples of heterocycloalkyls include: tetrahydrofuranyl, pyrrolidinyl, piperidinyl, 1,4-dioxanyl, morpholinyl, piperazinyl, oxyzolidinyl, decahydroquinolinyl.

When terms such as "optionally substituted" are used, these terms refer to all subsequent residues unless otherwise specified. That is, the term "optionally substituted alkyl, heteroalkyl, aryl, acyl" is to be read as "optionally substituted alkyl, optionally substituted heteroalkyl, optionally substituted aryl, optionally substituted acyl".

Depending on the substituents, in particular as a function of the substituent R, the compounds can be introduced into cells in a simple and directed manner. In one embodiment, R is preferably hydrogen.

In a further preferred embodiment, R represents a substrate, the z. B. enzymatically cleavable to convert prodrugs which have a cleavable product on the substituent R, in drugs.

That is, in a preferred embodiment, the substrate R is a cleavable product.

In a preferred embodiment, the substrate R is a cleavable substrate. This cleavable substrate is preferably one which, by proteolytic enzymes, plasmin, Cathepsin, cathepsin B, beta-glucuronidase, galactosidase, mannosidase, glucosidase, neuramidase, sucrose, maltase, fructosidase, glycosylases, prostate-specific antigen (PSA), urokinase-type plasminogen activator (u-PA), metalloproteinase, or an enzyme that can be specifically cleaved with the aid of targeted enzyme prodrug therapy, such as ADEPT, VDEPT, MDEPT, GDEPT, or PDEPT; or a substituent which can be cleaved or transformed under hypoxic conditions or by reduction by nitroreductase.

The radical R is preferably one selected from the group comprising monosaccharide, disaccharide or oligosaccharide, in particular hexoses, pentoses or heptoses, optionally as the deoxy derivative or amino derivative and optionally substituted by halogen, C _1-8 alkyl, C _{1 -8} acyl, C _1-8 heteroalkyl, C _3-7 cycloalkyl, C _3-7 heterocycloalkyl, C _4-12 aryl or C _4-12 heteroaryl, amino or amide groups, or with amino, amido or carboxyl moieties, the optionally substituted with halogen, C _1-8 alkyl, C _1-8 acyl, C _1-8 heteroalkyl, C _3-7 cycloalkyl, C _3-7 heterocycloalkyl, C _4-12 aryl or C _4-12 heteroaryl, amino - or amide radicals; Dextran, dipeptide, tripeptide, tetrapeptide, oligopeptide, peptidomimetics, or an antibody or antibody fragment, or combinations thereof.

With the aid of the substrate R, targeting of the compounds according to the invention to target structures is possible. D. h., It is a targeted coupling of the compounds of the invention to target structures and a corresponding introduction of these compounds of the invention in z. B. selected cells and cell types possible. The compounds of the invention wherein R is a substrate other than H, have a cleavable or transformable group. The cleavage or transformation of the compound of the invention with R except H can be carried out by chemical, photochemical, physical, biological or enzymatic processes under the appropriate conditions. These conditions include, for. B. the provision of appropriate enzymes, the change of the surrounding environment, the action of z. As radiation, such as UV light, etc. The skilled person are known methods. The substrate is suitably accessible to known methods such as ADEPT (Antibody Directed Enzyme Prodrug Therapy), PDEPT (Polymer-Directed Enzyme Prodrug Therapy) or MDEPT (Macromolecular-Directed Enzyme Prodrug Therapy), VDEPT (Virus-Directed Enzyme Prodrug Therapy) or GDEPT ( Gene-Directed Enzyme Prodrug Therapy).

wobei R unabhängig voneinander Wasserstoff oder CH₃ ist und n ist eine ganze Zahl aus 3, 4 oder 5.Particularly preferred compounds are the compounds shown below:

wherein R is independently hydrogen or CH ₃ and n is an integer of 3, 4 or 5.

In a further preferred embodiment of the present invention, the components A and B are identical radicals. According to the invention, hybrid compounds are provided from structures I and II, but the compounds are preferably those in which the structures A and B are identical radicals, as in the particularly preferred compounds III and IV, as shown above.

Furthermore, the present application is directed to pharmaceutical compositions containing the compounds of the invention, optionally with pharmaceutically acceptable carriers or diluents. The compounds according to the invention can be present in the form of pharmaceutically acceptable salts or solvates.

Pharmaceutically acceptable salts are in particular acid addition salts, which are formed according to amine groups. Likewise, base addition salts are possible or corresponding zwitter addition salts.

The term "pharmaceutically acceptable solvates" refers to the association of one or more solvent molecules and a compound of the invention. Examples of such solvent molecules that form pharmaceutically acceptable solvates include water, isopropyl alcohol, ethanol, methanol, DSMO, ethyl acetate, and acetic acid.

The compounds of the invention are particularly suitable for the preparation of pharmaceutical compositions suitable in tumor therapy. Monomers of the bifunctional compounds of the invention are known as cytotoxic compounds suitable for tumor therapy. The present invention includes pharmaceutical compositions which contain the compounds according to the invention in addition to the customary carriers or diluents. The preparation of the pharmaceutical preparations listed above is carried out in the usual manner by known methods, for. B. by mixing the or the active ingredients or carriers.

In general, the compounds of the invention may be administered in total amounts of from 0.5 to about 500, preferably 1 to 150 mg / kg of body weight per 24 hours, optionally in the form of several single doses, to achieve the desired results. The person skilled in the art knows the possibilities for determining the dose amount. This can be done depending on the age, the body weight, the type and severity of the disease of the patient, the method of preparation and the application of the drug and the period or the interval of administration.

Description of the pictures

In the 1 describes the synthesis of the β-D-galactosides of the bifunctional (1S) -seco-CBI and the (1S, 10R) -anti-methyl-seco-CBI derivatives, as well as the corresponding seco-drugs: Synthesis of the β-D Galactosides of the bifunctional (1S) -seco-CBI and (1S, 10R) -anti-methyl-seco-CBI derivatives and the corresponding seco-drugs: a) 87, BF ₃ · OEt ₂ , CH ₂ Cl ₂ , MS (4 Å), -10 ° C, 3.5 h, then BF ₃ · OEt ₂ , RT, 5.5 h; b) 66a-c (n = 3-5), NEt ₃ , DMF, RT, 20h; c) Preparative HPLC: Kromasil ^® 100 C18 (250 20 mm, 7 microns), A = H ₂ O, B = MeOH, gradient: A / B = 30:70 → 0: 100 in 6.5 min; d) NaOMe / MeOH, RT, 2h; e) Preparative HPLC: Kromasil ^® 100 C18 (250 20 mm, 7 microns), A = H ₂ O, B = MeOH, gradient: A / B = 70:30 → 0: 100 in 15 min; f) 4N HCl / EtOAc, RT, 3h; g) 66a-c (n = 3-5), pyridine, DMF, RT, 20 h; h) Preparative HPLC: Kromasil ^® 100 C18 (250 20 mm, 7 microns), A = H ₂ O, B = MeOH, gradient: A / B = 30:70 → 0: 100 in 6.5 min.

The 2 shows the racemic synthesis route for the preparation of N-Boc-seco-CBI (rac-69) with subsequent separation of the enantiomers via HPLC on a chiral stationary phase.

The 3 shows the asymmetric synthetic route to represent (1S) -N-Boc-seco-CBI ((-) - (1S) -69).

The 4 shows the in vitro cytotoxicity determined in the A549 cell line of the bifunctional (1S, 10R) -anti-methyl-seco-CBI derivatives with n = 3 (o.E. = without enzyme, m.E. = with enzyme) ,

The 5 shows the in vitro cytotoxicity determined in the A549 cell line of the bifunctional (1S, 10R) -anti-methyl-seco-CBI derivatives with n = 4 (o.E. = without enzyme, m.E. = with enzyme) ,

The 6 shows the in vitro cytotoxicity determined in the A549 cell line of the bifunctional anti-methyl seco-CBI derivatives with n = 5 (o.E. = without enzyme, m.M. = with enzyme).

In the following, the invention will be explained in more detail with the aid of the examples, without the invention being restricted to these.

Examples

Materials:

The cell lines used are available via ATCC and were cultured in the recommended media.
Culture medium for A549: DMEM (Dulbecco's Modified Eagles Medium) with 4.5 g / l glucose (Biochrom, T043-10). The medium was supplemented with 4 mM L-glutamine and 3.7 g / l sodium bicarbonate.
Medium additives: 10% FBS (fetal calf serum) from Biochrom, inactivated for 30 min at 56 ° C.
Enzyme: β-D-galactosidase (EC 3.2.1.23) from Escherichia coli G 5635 (Sigma), activity: 250-600 units (units (U)) per mg protein at pH 7.3 and 37 ° C, 1 U = 1 μmol Substrate turnover per minute.
PBS buffer: solution of the PBS-dry substance (Phosphate Buffered Saline) from Biochrom KG in bidistilled water. Salt concentrations in the buffer solution with pH 7.4: 137.0 mM NaCl, 2.7 mM KCl, 8.1 mM Na ₂ HPO ₄ , 1.1 mM KH ₂ PO ₄ .
Rotational values: Polarimeter Model 241 from Perkin-Elmer.
Infrared Spectra: Model Vector 22 from Bruker. Solids were measured as KBr pellets, liquids as a film between KBr pellets. For calibration, the polystyrene band served at 1601 cm ^-1 .
UV / VIS spectra: Lambda 2 model from Perkin-Elmer.
¹ H NMR spectra: Models Mercury-200 (200 MHz), Mercury-300 and Unity-300 (each 300 MHz) and Unity Inova-600 (600 MHz). the company Varian. The chemical shifts are given in units of the δ scale. Tetramethylsilane (δ _TMS = 0.00 ppm) or the indicated solvent served as an internal standard. The following abbreviations are used to denote the multiplicities of the signals: s (singlet), d (doublet), t (triplet), q (quartet), m (multiplet), m _c (centered multiplet), br (broad signal). The following abbreviations are used to refer to aromatic protons or carbon atoms: i (ipso), o (ortho), m (meta), p (para). The spectra were usually interpreted according to the first order. The coupling constants J are given in Hertz (Hz).
¹³ C NMR spectra: Models Mercury-200 (50 MHz), Mercury-300 and Unity-300 (each 75 MHz) and Unity Inova-600 (150 MHz). The internal standard was tetramethylsilane or the specified solvent. The chemical shifts are taken from the ¹ H broadband decoupled spectra, the multiplicities of the signals were determined in multiple selection experiments (APT pulse sequence).
Mass spectra: The EI spectra were recorded using a Finnigan double-focusing sector field mass spectrometer MAT 95. ESI spectra were recorded using a Finnigan LCQ ion mass spectrometer LCQ. The quotients from mass to charge are given as well as the relative intensities in parentheses relative to the base peak (I = 100). The measurement of the ESI-HRMS spectra was carried out on a Bruker Bruker APK Tkerla Fourier Transform Ion Cyclotron Resonance (FTICR) mass spectrometer, which was equipped with an Bruker Apollo ESI source and a 74900 series syringe pump from Cole Corporation. Parmer is equipped. The flow of the syringe pump was 2 μL / min. To record the spectra and to evaluate the program XMASS Bruker.
Thin layer chromatography (TLC): DC ready-made films SIL G / UV 254 from Macherey-Nagel & Co. (layer thickness 0.25 mm) were used. Indicated are R _f values (running height relative to the solvent front). Abbreviations for the solvents used are: DMSO (dimethylsulfoxide), EtOAc (ethyl acetate), CH ₂ Cl ₂ (dichloromethane), MeOH (methanol). In addition to the UV detection, a vanillin-sulfuric acid solution (0.5 g vanillin, 3 mL concentrated sulfuric acid, 85 mL methanol and 10 mL acetic acid) and a solution of molybdophosphoric acid (5% in methanol) served as staining reagents.
Column chromatography: All column chromatographic separations were carried out with silica gel 60 (particle size: 0.032-0.063 mm) from Merck.
High pressure liquid chromatography (HPLC):
Analytical HPLC: Analytical separations except for the oligonucleotide experiments were carried out on a Jasco HPLC system equipped with a solvent pump PU-2080, a mixing chamber LG-1590-04, a multi-wavelength detector MD-2010 Plus and the controller LC-Net II / ADC, made. In addition, an automatic sample changer (autosampler AS-2055) from the same company was connected. The computer programs Borwin PDA, HSS 2000 and Borwin Chromatography from Jasco were used for operation, data acquisition and data evaluation. For the measurements, the finished analytical column Chiralcel ^® OD (250 x 4.6 mm, particle size: 10 .mu.m) Daicel Chemical Industries Ltd. and Kromasil 100 C18 (5 μm, 250 × 4 mm) from Jasco. N-hexane, dichloromethane, and isopropanol were used as solvents of HPLC grade (Chiralcel ^® OD) and bidistilled water (addition of 0.1 vol .-% trifluoroacetic acid for peptide synthesis or 12:06 vol .-% of concentrated hydrochloric acid) and acetonitrile or HPLC grade methanol (Kromasil 100 C18). All samples were membrane filtered and the solvents degassed.
Preparative HPLC: Preparative separations were carried out on a Jasco HPLC system equipped with two solvent pumps model PU-2087 PLUS and a UV detector model UV-2075 PLUS. A finishing column Chiralpak ^® IA were used (250 x 20 mm, particle size: 5 .mu.m) and a prepacked column of the type Kromasil 100 C18 (7 .mu.m, 250 x 20 mm) from Jasco 100 C18 (in conjunction with a guard column of the type Kromasil 5 μm, 50 × 20 mm) from Jasco. N-heptane, n-hexane and dichloromethane were used as solvents of HPLC grade (Chiralpak ^® IA) and bidistilled water (addition of 0.1 vol .-% trifluoroacetic acid for peptide synthesis) and acetonitrile or methanol in HPLC grade (Kromasil 100 C18) , All samples were membrane filtered and the solvents degassed.

example 1

Racemic synthesis of the seco-CBI backbone, as shown in FIG.

A. (E / Z) -2-amino-4-benzyloxy-N- (tert-butyloxycarbonyl) -N- (3-chloro-2-propenyl) -1-iodo-naphthalene (75)

To a suspension of NaH (420 mg, 10.5 mmol, 2.5 eq., 60% suspension in mineral oil) in absolute DMF (30 mL) was added via transfer cannula a solution of the amide 73 (2.00 g, 4.21 mmol, 1.0 eq absolute DMF (20 mL). The mixture was stirred at room temperature for 2 h, (E / Z) -1,3-dichloropropene (74) (870 μL, 8.42 mmol, 2.0 eq.) Was added and the mixture was stirred for a further 6 h at room temperature. The reaction was quenched by careful addition of a saturated NH ₄ Cl solution (50 mL) and the reaction mixture was treated with ethyl acetate (100 mL) and water (50 mL). It was then extracted with ethyl acetate (3 × 150 ml), the combined organic phases were washed with water (4 × 200 ml) and saturated NaCl solution (200 ml _), dried over MgSO ₄ and the solvent was removed in vacuo. Silica gel column chromatography (n-pentane / EtOAc = 10: 1) provided the title compound 75 as a yellow oil (2.29 g, 4.16 mmol, 99%).
R _f = 0.51, 0.44 (n-pentane / EtOAc = 10: 1).
¹ H-NMR (200 MHz, CDCl ₃ ): δ = 1.33 / 1.58 (each s, total 9 H, C (CH ₃ ) ₃ ), 3.72-4.34 (m, 1H, 1'-H _a ), 4.47 -4.63 (m, 1H, 1'-H _b), 5.26 (s _br, 2H, OC H ₂ Ph), 5.93-6.18 (m, 2H, 2'-H, 3'-H), 6.65-6.79 ( m, 1H, 3-H), 7.31-7.63 (m, 7H, 6-H, 7-H, 5xPh-H), 8.10, 8.22 (each d, J = 8.0 Hz, each 1H, 5-H, 8) -H).
¹³ C-NMR (50 MHz, CDCl ₃ ): δ = 28.3 / 28.5 (C ( C H ₃ ) ₃ ), 46.0 / 49.1 (C-1 '), 70.3 / 70.4 (O C H ₂ Ph), 80.7 ( C (CH ₃ ) ₃ ), 95.0 / 95.1 (C-1), 107.1 / 107.7 (C-3), 120.6 / 120.8 (C-3 '), 122.5 (C-5), 125.5 (C-4a), 126.3, 126.4, 127.3 (C-6, C-2 ', Ph-C _o), 128.1 / 128.2 (Ph-C _p), 128.5 / 128.6 (C-8), 128.7 (Ph-C _m), 132.7 ( C-7), 135.3 (C-8a), 136.3 / 136.4 (Ph-C _i ), 142.4 / 142.8 (C-2), 153.7 / 153.9 (C = O), 155.3 (C-4).

B. rac-5-Benzyloxy-3- (tert-butyloxycarbonyl) -1-chloromethyl-1,2-dihydro-3H-benz [e] indole (rac-58)

The iodoaromatic 75 (2.29 g, 4.16 mmol, 1.0 eq.) Was dissolved in toluene (50 mL) and the solution was degassed thoroughly by passing in an argon gas stream. Tris (trimethylsilyl) silane (1.46 mL, 4.58 mmol, 1.1 eq.) And AIBN (171 mg, 1.04 mmol, 0.25 eq.) Were added and the mixture was stirred at 80 ° C. for 4 h. After cooling, the reaction mixture was admixed with a 10% strength aqueous KF solution (50 ml) and stirred at room temperature for 1 h. The organic phase was dried over MgSO ₄ , the solvent removed in vacuo and the residue purified by column chromatography on silica gel (gradient: n-pentane / EtOAc = 50: 1 → 20: 1). The product rac-58 was obtained as a white solid (1.38 g, 3.26 mmol, 78%).
R _f = 0.50 (n-pentane / EtOAc = 10: 1).
¹ H-NMR (300 MHz, CDCl ₃ ): δ = 1.62 (s, 9H, C (CH ₃ ) ₃ ), 3.44 (t, J = 10.9 Hz, 1H, 10-H _a ), 3.92-4.01 (m , 2H, 1-H, 10-H _b ), 4.13 (t, J = 10.7 Hz, 1H, 2-H _a ), 4.28 (d, J = 11.1 Hz, 1H, 2-H _b ), 5.27 (s , 2H, OC H ₂ Ph), 7.31-7.56 (m, 7H, 7-H, 8-H, 5xPh-H), 7.65 (d, J = 8.4 Hz, 1H, 9-H), 7.87 (s _br , 1H, 4-H), 8.30 (d, J = 8.4 Hz, 1H, 6-H).
¹³ C-NMR (50 MHz, CDCl _3): δ = 28.5 (C (C H _{3) 3),} 41.6 (C-1), 46.5 (C-10), 53.0 (C-2), 70.3 (O C H ₂ Ph), 81.2 ( C (CH ₃ ) ₃ ), 96.4 (C-4), 114.0 (C-9b), 122.4 (C-5a), 121.7, 123.1, 123.6 (C-6, C-7, C-9), 127.5 (Ph-C _o ), 127.6 (C-8), 128.0 (Ph-C _p ), 128.5 (Ph-C _m ), 130.2 (C-9a), 136.7 (Ph-C _i ) , 143.4 (C-3a), 152.6 (C = O), 156.0 (C-5).

C. rac-3- (tert -butyloxycarbonyl) -1-chloromethyl-5-hydroxy-1,2-dihydro-3H-benz [e] indole (rac 69)

To a solution of the benzyl ether rac-58 (200 mg, 472 μmol, 1.0 eq.) In freshly distilled THF at 35 ° C first palladium on activated carbon (10%, 100 mg, 94 .mu.mol, 0.2 eq. Regarding Pd) and then ammonium formate (25% aqueous solution, 1.0 mL). After stirring at 30 ° C for 25 min, it was immediately filtered from the solid through a pad of celite, rinsing thoroughly with acetone (400 mL). The solvent was removed in vacuo, and after column chromatography on silica gel (n-pentane / EtOAc = 5: 1), 147 mg of an indissoluble mixture (83:17) from the title compound rac-58 (124 mg, 372 μmol, 79%) and the dechlorinated racemic by-product (23 mg, 75.5 μmol, 16%) were obtained.
R _f = 0.54 (n-pentane / EtOAc = 5: 1). C ₁₈ H ₂₀ CINO ₃ (333.82). ber .: 334.1205 Found: 334.1205, [M + H] ⁺ (ESI-HRMS).

D. Separation of enantiomers of rac-3- (tert-butyloxycarbonyl) -1-chloromethyl-5-hydroxy-1,2-dihydro-3H-benz [e] indole (rac-69)

rac-69 / iso-propanol was dissolved in a mixture of n-hexane (1: 1) (c = 25 mg / mL) and 1 mL of this solution into the preparative HPLC system (column: Chiralpak ^® IA, 250 x 20 mm, particle size: 5 μm, mobile phase: n-hexane / isopropanol = 97: 3, flow: 18 mL / min, pressure: 5.7 MPa). Fractional capture of the eluate (UV detector: λ = 255 nm) yielded the enantiomers (+) - (1R) -69 and (-) - (1S) -69. Analytical data for (+) - (1R) -69: HPLC (preparative): t _R : 11.9 min Fraction: 11.1-13.0 min HPLC (analytical): Pillar: Chiralcel OD, 250 × 4.6 mm, 10 μm eluent: 2% iso-propanol in n-hexane River: 0.8 mL / min t _R : 16.4 min, 99.9% ee [Α] 20 / D = + 48.6 ° (c = 0.28, CHCl ₃ ). Analytical data for (-) - (1S) -69: HPLC (preparative): t _R : 14.0 min Fraction: 13.6-15.8 min HPLC (analytical): Pillar: Chiralcel® OD, 250 × 4.6 mm, 10 μm eluent 2% iso-propanol in n-hexane River: 0.8 mL / min t _R : 19.3 min, 99.9% ee [Α] 20 / D = -45.8 ° (c = 0.24 CHCl _3).
UV (CH ₃ CN): λ _max (Ig ε) = 208.5 nm (4,236), 220.0 (4,216), 255,0 (4,853), 303.5 (3,933), 314.5 (3,992), 342.0 (3,489).
IR (KBr): ṽ (cm ^-1 ) = 3368, 2977, 1682, 1629, 1585, 1522, 1480, 1413, 1339, 1236, 1143, 1058, 911, 852, 758, 674.
¹ H-NMR (300 MHz, CDCl ₃ ): δ = 1.62 (s, 9H, C (CH ₃ ) ₃ ), 3.41 (t, J = 11.4 Hz, 10-H _a ), 3.90-3.98 (m, 2H , 1-H, 10-H _b ), 4.12 (t, J = 10.5 Hz, 1H, 2-H _a ), 4.25 (d, J = 11.5 Hz, 1H, 2-H _b ), 7.33 (dt, J = 7.6, 1.0 Hz, 1H, 7-H), 7.49 (dt, J = 7.6, 1.0 Hz, 1H, 8-H), 7.62 (d, J = 8.4 Hz, 1H, 9-H), 7.82 (s _br , 1H, 4-H), 8.21 (d, J = 8.4 Hz, 1H, 6-H).
¹³ C-NMR (125 MHz, CDCl _3): δ = 28.5 (C (C H _{3) 3),} 41.8 (C-1), 46.5 (C-10), 53.1 (C-2), 81.8 (C ( CH ₃ ) ₃ ), 99.1 (C-4), 114.3 (C-9b *), 121.6 (C-5 *), 121.7 (C-9), 122.9 (C-7), 123.6 (C-6), 127.6 (C-8), 130.4 (C-9a), 141.1 (C-3a), 153.3 (C = O), 154.1 (C-5).
MS (ESI): m / z (%) = 334.1 (15) [M + H] ⁺ , 278.1 (100) [MC ₄ H ₈ + H] ⁺ . C ₁₈ H ₂₀ ClNO ₃ (333.82). ber .: 334.1205 Found: 334.1205, [M + H] ⁺ (ESI-HRMS).

Example 2

Asymmetric synthesis of the (1S) -seco-CBI backbone as shown in FIG.

A. (+) - (2S) -glycidyl-3-nitrobenzenesulfonate ((+) - (S) -77)

A solution of (+) - (R) -glycidol (76) (4.85 g, 65.5 mmol, 1.0 eq.) And triethylamine (10.9 mL, 7.95 g, 78.6 mmol, 1.2 eq.) In absolute toluene (130 mL) Chilled to 0 ° C and treated portionwise with m-nitrobenzenesulfonyl chloride (14.5 g, 65.5 mmol, 1.0 eq.). After warming to room temperature, the mixture was stirred for 1 h at this temperature, the mixture was then filtered through Celite and a little silica gel and washed with toluene (250 mL). After removal of the solvent in vacuo, the crude product was dissolved in ethanol at 40 ° C. (maximum 20 mL / g) and crystallized by slow cooling to -30 ° C. The target compound (+) - (S) -77 (7.70 g, 29.7 mmol, 45%, (R) -enantiomer undetectable) was obtained as colorless needles. Analytical data for (-) - (R) -77: HPLC (analytical): Pillar: Chiralpak ^® IA, 250 x 4.6 mm, 5 micron eluent 14% THF in n-hexane River: 0.8 mL / min t _R : 52.3 min Analytical data for (+) - (S) -77: HPLC (analytical): Pillar: Chiralpak ^® IA, 250 x 4.6 mm, 5 micron eluent 14% THF in n-hexane River: 0.8 mL / min t _R : 48.4 min R _f = 0.67 (Et ₂ O).
[Α] 20 / D = + 22.7 ° (c = 2.14, CHCl ₃ ).
¹ H-NMR (300 MHz, CDCl ₃ ): δ = 2.63 (dd, J = 4.7, 2.5 Hz, 1H, 3'-H _a ), 2.85 (m _c , 1H, 3'-H _b ), 3.23 ( m _c , 1H, 2'-H), 4.05 (dd, J = 11.7, 6.6 Hz, 1H, 1'-H _a ), 4.50 (dd, J = 11.6, 2.9 Hz, 1H, 1'-H _b ) , 7.83 (t, J = 8.1 Hz, 1H, 5-H), 8.27 (ddd, J = 7.8, 1.9, 1.2 Hz, 1H, 6-H), 8.54 (ddd, J = 8.1, 2.2, 1.1 Hz, 1H, 4-H), 8.78 (m _c, 1H, 2-H). ¹³ C-NMR (75 MHz, CDCl _3): δ = 44.5 (C-3 '), 48.7 (C-2'), 71.7 (C-1 '), 123.2 (C-2), 128.4 (C-4 ), 130.8 (C-5), 133.3 (C-6), 138.0 (C-1), 148.3 (C-3).

B. (+) - {(2'R) -2-amino-4-benzyloxy-N- (tert-butyloxycarbonyl) -N- (2,3-epoxypropyl) -1-iodo-naphthalene} ((+) ( 2'R) -78)

Amide 73 (4.76 g, 10.0 mmol, 1.0 eq.) Was dissolved in DMF (125 mL) and heated to 40 ° C. NaH (721 mg, 30.1 mmol, 3.0 eq, 60% suspension in mineral oil) and the nosylate (+) - (S) -77 (3.90 g, 15.0 mmol, 1.5 eq.) Were added after addition of the mixture for 3 Stirred at 40 ° C h. After cooling to room temperature, saturated NH ₄ Cl solution (250 mL) was added to the suspension, extracted with Et ₂ O (3 × 200 mL), and the combined organic phases were washed with water (200 mL) and saturated NaCl (2 × 200 mL), dried over Na ₂ SO ₄ and the solvent removed in vacuo. Column chromatographic purification on silica gel (n-pentane / EtOAc = 5: 1) gave the product (+) - (2'R) -78 (5.08 g, 9.56 mmol, 96%) as a mixture of atropisomers as a white solid.
R _f = 0.53 (n-pentane / EtOAc = 7: 3).
[Α] 20 / D = + 8.0 ° (c = 0.6, CHCl ₃ )
UV (CH ₃ CN): λ _max (Ig ε) = 216.0 nm (4,682), 243.5 (4,437), 304.0 (3,948).
IR (KBr): ṽ (cm ^-1 ) = 3063, 2978, 2929, 1701, 1617, 1591, 1562, 1503, 1454, 1404, 1370, 1326, 1261, 1231, 1156, 1102, 1029, 973, 912, 879, 841, 760, 736, 697, 642.
¹ H-NMR (300 MHz, tetrachloroethane-d ₂ , 100 ° C, the respective signals of the two atropisomers are denoted by (a) and (b)): δ = 1.46 (s, 18H, 2xC (CH ₃ ) ₃ ) , 2.46 (dt, J = 4.7, 2.6 Hz, 2H, 3'-H _a (a), 3'-H _a (b)), 2.73 (dt, J = 4.8, 9.0 Hz, 2H, 3'-H _b (a), 3'-H _b (b)), 3:12 to 3:19 (m, 1H, 2'-H (a)), 3.28 (dd, J = 14.4, 6.6 Hz, 1H, 1'-H _a (a)), 3.35-3.42 (m, 1H, 2'-H (b)); 3:49 (dd, J = 14.7, 4.9 Hz, 1H, 1'-H _a (b)), 4:09 to 4:17 (m, 2H, 1'-H _b (a), 1'-H _b (b)), 5.32, 5.33 (2xs, 4H, OC H ₂ Ph (a), OC H ₂ Ph (b)), 6.89, 6.99 (2xs, 2H, 3-H (A) 3-H (b)), 7.34-7.67 (m , 14H, 6-H (a), 6-H (b), 7-H (a), 7-H (b), 5xPh-H (a) 5xPh-H (b)), 8.25 (m _c , 2H, 8-H (a), 8-H (b)), 8:36 (m _c, 2H, 5-H (a) and 5-H (b)
¹³ C-NMR (75 MHz, tetrachloroethane-d ₂ , 100 ° C, the respective signals of the two atropisomers are denoted by (a) and (b)): δ = 29.8 (2 signals) (2xC ( C H ₃ ) ₃ ), 47.3, 47.5 (C-3 '(a), C-3' (b)), 51.0 (C-2 '(a)), 51.5 (C-2' (b)), 52.9 (C-1 '(b)), 54.5 (C-1' (a)), 72.2, 72.3 (2xO C H ₂ Ph), 82.1 (2 signals) (2x C (CH ₃ ) ₃ ), 96.2, 96.3 (C-1 (a), C-1 (b)), 109.6, 109.7 (C-3 (a), C-3 (b)), 123.9 (C-5 (a), C-5 (b)), 127.2 ( C-4a (a), C-4a (b)), 127.7 (2 signals), 128.6, 128.8, 129.4, 129.8, 129.9, 130.0 (2 signals) (C-6 (a), C-6 (b) , C-7 (a), C-7 (b), C-8 (a), C-8 (b), 2x Ph-C _o (a), 2x Ph-C _o (b), 2x Ph-C _m ( a), 2x Ph-C _m (b), Ph-C _p (a), Ph-C _p (a)), 136.9 (2 signals) (C-8a (a), C-8a (b)), 137.9 , 138.0 (Ph-C _i (a), Ph-C _i (b)), 144.8, 145.3 (C-2 (a), C-2 (b)), 155.0, 155.2 (C = O (a), C = O (b)), 157.0, 157.1 (C-4 (a), C-4 (b)).
MS (ESI): m / z (%) = 1085.2 (100) [2M + Na] ⁺ , 554.1 (97) [M + Na] ⁺ . C ₂₅ H ₂₆ INO ₄ (531.38). about: 554.0799 Found: 554.0802, [M + Na] ⁺ (ESI-HRMS).

C. (+) - {(1S) -5-Benzyloxy-3- (tert-butyloxycarbonyl) -1-hydroxymethyl-1,2-dihydro-3H-benz [e] indole} ((+) - (1S) - 79)

A solution of freshly sublimed ZnCl ₂ (1.36 g, 9.96 mmol, 2.11 eq.) In absolute THF (80 mL) was cooled to 0 ° C, added slowly with methyl lithium (24.9 mL of a 1.6 M solution in Et ₂ O, 39.8 mmol, 8.42 eq.) And stirred for 30 min at this temperature. The solution was then cooled to -78 ° C., admixed dropwise with TMS-NCS (1.42 mL, 1.31 g, 9.96 mmol, 2.11 eq.), Warmed again to 0 ° C. and stirred for 30 min at this temperature. After cooling again to -78 ° C, to the reaction mixture was added dropwise a solution of the epoxide ((+) - (2'R) -78 (2.51 g, 4.73 mmol, 1.0 eq.) In absolute THF (80 mL) for 30 min After warming to 0 ° C., the reaction solution was first stirred for 1.5 h at this temperature, then warmed to room temperature and stirred for a further 12 h, after which the mixture was carefully washed with saturated NH ₄ Cl solution (250 mL extracted, extracted with CH ₂ Cl ₂ (4 × 250 mL), the combined organic phases dried over Na ₂ SO ₄ and the solvent removed in vacuo, purified by column chromatography on silica gel (n-pentane / EtOAc = 4: 1) Target compound (+) - (1S) -79 (1.12 g, 2.76 mmol, 58%, 96.1% ee) as a white solid Analytical Data for (-) - (1R) -79: HPLC (analytical): Pillar: Chiralcel ^® OD, 250 x 4.6 mm, 10 micron eluent: 5% iso-propanol in n-hexane River: 0.8 mL / min t _R : 13.6 min Analytical data for (+) - (1S) -79: HPLC (analytical): Pillar: Chiralcel ^® OD, 250 x 4.6 mm, 10 micron eluent: 5% iso-propanol in n-hexane River: 0.8 mL / min t _R : 14.3 min R _f = 0.35 (n-pentane / EtOAc = 3: 1).
[Α] 20 / D = + 4.3 ° (c = 1.79, CH ₂ Cl ₂ ).
UV (CH ₃ CN): λ _max (Ig ε) = 207.5 nm (4,447), 217.5 (4,396) 255.0 (4,834) 304.0 (3,994) 315.0 (4,072) 341.0 (3,550).
IR (KBr): ṽ (cm ^-1 ) = 3446, 2975, 1699, 1626, 1582, 1517, 1460, 1407, 1332, 1268, 1142, 1032, 906, 845, 760, 696,
¹ H-NMR (300 MHz, CDCl ₃ ): δ = 1.59 (s, 9H, C (CH ₃ ) ₃ ), 3.72-3.87 (m, 2H, 1-H, 10-H _a ), 3.91-3.97 ( m, 1H, 10-H _b ), 4.08-4.15 (m, 1H, 2-H _a ), 4.19-4.23 (m, 1H, 2-H _b ), 5.26 (s _br , 2H, OC H ₂ Ph) , 7.28-7.55 (m, 7H, 7-H, 8-H, 5xPh-H), 7.70 (d, J = 8.4 Hz, 1H, 9-H), 7.89 (s _br , 1H, 4-H), 8.28 (d, J = 8.2 Hz, 1H, 6-H).
¹³ C-NMR (125 MHz, CDCl _3): δ = 28.5 (C (C H _{3) 3),} 41.3 (C-1), 52.4 (C-2), 64.7 (C-10), 70.3 (O C H ₂ Ph), 80.7 ( C (CH ₃ ) ₃ ), 96.5 (C-4), 114.4 (C-9b), 122.3 (C-5a), 122.2 (C-9), 122.9 (C-7), 123.4 (C-6), 127.3 (C-8), 127.5, 127.9, 128.5 (5x Ph-CH), 130.6 (C-9a), 136.9 (Ph-C _i ), 141.9 (C-3a), 152.7 (C = O), 155.6 (C-5).
MS (ESI): m / z (%) = 350.1 (100) [MC ₄ H ₈ + H] ⁺ , 406.2 (44) [M + H] ⁺ , 428.2 (22) [M + Na] ⁺ , 828.4 ( 24) [M + NH ₄ ] ⁺ . C ₂₅ H ₂₇ NO ₄ (405.49). calculated: 406.2013 Found: 406.2013, [M + H] ⁺ (ESI-HRMS).

D. (-) - {(1S) -3- (tert-Butyloxycarbonyl) -1-chloromethyl-5-hydroxy-1,2-dihydro-3H-benz [e] indole} ((-) (1S) 58)

A solution of (+) - (1S) -58 (2.15 g, 5.30 mmol, 1.0 eq.) In absolute CH ₂ Cl ₂ (20 mL) was treated at room temperature with triphenylphosphine (4.17 g, 15.9 mmol, 3.0 eq.) And CCl ₄ (4.60 mL, 7.34 g, 47.7 mmol, 9.0 eq.) And stirred for 3 h at this temperature. Removal of the solvent in vacuo followed by purification by column chromatography on silica gel (n-pentane / EtOAc = 20: 1) gave the title compound (-) (1S) -58 (2.13 g, 5.03 mmol, 95%, 96.1% ee) as white Obtained solid. Analytical data for (+) - (1R) -58: HPLC (analytical): Pillar: Chiralcel ^® OD, 250 x 4.6 mm, 10 micron eluent: 2% iso-propanol in n-hexane River: 0.8 mL / min t _R : 9.6 min Analytical data for (-) - (1S) -58: HPLC (analytical): Pillar: Chiralcel ^® OD, 250 x 4.6 mm, 10 micron eluent: 2% iso-propanol in n-hexane River: 0.8 mL / min t _R : 13.4 min

The further steps were carried out as in Example 1, step C., to give the compound enantiomeric -3- (tert-butyloxycarbonyl) -1-chloromethyl-5-hydroxy-1,2-dihydro-3H-benz [e] indole (rac 69) to obtain compounds.

Example 3

Synthesis of the bifunctional seco-CBI derivatives and the corresponding β-D-galactosides

A. (-) - {1,5-bis [(1S) -1-chloromethyl-5-hydroxy-1,2-dihydro-3H-benz [e] indol-3-yl] -pentane-1,5-dione ]} ((-) - (1'S) -55a, n = 3)

The phenol (-) - (1S) -69 (61.0 mg, 183 μmol, 2.0 eq.) Was dissolved in 4 M HCl / ethyl acetate (4.5 mL) and, after stirring at room temperature for 1 h, the solvent was removed in vacuo. The residue was dried in vacuo for 1 h, dissolved in DMF (5 mL), the solution was cooled to 0 ° C. and treated with glutaric acid chloride (66a) (12.0 μL, 15.4 mg, 91.4 μmol, 1.0 eq.) And pyridine (29.5 μL, 28.9 mg, 365 μmol, 4.0 eq.). After stirring for 20 h at room temperature, the reaction mixture was diluted with DMF (5 mL) and purified in successive injections (0.4 mL each) by preparative HPLC. The product (-) - (1'S) -55a (38.9 mg, 69.0 μmol, 76%) was obtained as a white solid. HPLC (preparative): Pillar: Kromasil 100 C18, 250 × 20 mm, 7 μm Guard column: - Gradient: Time [min] H ₂ O / MeOH 0 30/70 0-6.0 30/70 → 0/100 6.0-8.5 0/100 8.5-9.5 0/100 → 30/70 9.5-15 30/70 River: 16 mL min ^-1 λ: 254 nm t _R : 8.4 min [Α] 20 / D = -32.7 ° (c = 0.257, DMSO).
UV (CH ₃ CN): λ _max (lg ε) = 205.5 nm (4,550), 2,225 (4,406), 256.5 (4,909), 264.5 (4,921), 319.0 (4,332).
IR (KBr): ṽ (cm ^-1 ) = 3122, 2926, 1633, 1584, 1522, 1459, 1426, 1394, 1253, 1131, 1075, 1024, 857, 756, 717.
¹ H-NMR (600 MHz, DMSO-d ₆ ): δ = 1.98 (m _c , 2H, 3-H ₂ ), 2.57-2.62 (m, 2H, 2-H _a , 4-H _a ), 2.68- 2.74 (m, 2H, 2H _b, 4-H _b), 3.79 (dd, J = 10.9, 8.3 Hz, 2H, 2x10'-H _a), 3.99 (dd, J = 10.9, 3.2 Hz, 2H, 2x10'-H _b ), 4.14-4.20 (m, 4H, 2x1'-H, 2x2'-H _a ), 4.34 (m _c , 2H, 2x2'-H _b ), 7.32 (m _c , 2H, 2x7 ' -H), (m _c 7:49, H 2H-2x8',) 7.78 (d, J = 8.3 Hz, 2H, H-2x9') 8.02 (s, 2H, H-2x4') eight nine (d, J = 8.4 Hz, 2H, 2x6'-H), 10.31 (s, 2H, 2xOH).
¹³ C-NMR (125 MHz, DMSO-d ₆ ): δ = 19.3 (C-3), 34.4 (C-2, C-4), 40.7 (2 × C-1 '), 47.6 (2 × C-10'), 52.6 (2xC-2 '), 99.7 (2xC-4'), 113.6 (2xC-5a '), 121.5 (2xC-9b'), 122.3 (2xC-9 '), 122.4 (2xC-7'), 122.9 ( 2xC-6 '), 127.0 (2xC-8'), 129.8 (2xC-9a '), 141.8 (2xC-3a'), 154.0 (2xC-5 '), 170.4 (2xC = O).
MS (ESI): m / z (%) = 1125.4 (8) [2M-H] ^- , 561.2 (96) [MH] ^- , 525.2 (100) [M-2H-Cl] ^- . C ₃₁ H ₂₈ Cl ₂ N ₂ O ₄ (563.47). ber .: 585.1318 Found: 585.1317, [M + Na] ⁺ (ESI-HRMS).

Example 4

(-) - {1,5-bis [(1S) -1-chloromethyl-5- (β-D-galactopyranosyl) -1,2-dihydro-3H-benz [e] indol-3-yl] pentan-1 , 5-dione]} ((-) - (1'S) -53a, n = 3)

The acetyl protected galactoside (1'S) -71a (148 mg, 121 μmol, 1.0 eq.) Was suspended in absolute methanol (35 mL) and washed at 0 ° C with NaOMe in methanol (90 μL of a 5.4 M solution, 484 μmol, 4.0 eq .). After warming to room temperature and stirring for 30 minutes at this temperature, the substrate had completely dissolved. Stirring was continued for a further 1.5 h at room temperature, the reaction solution followed by water (3.0 mL) and as long as acidic ion exchanger added (Amberlite IR-120 ^®), until the solution was neutral (pH = 7). The mixture was separated via transfer cannula from the ion exchanger, the same washed with methanol (2 × 6 mL), the organic phases combined and the solvent removed in vacuo. The resulting residue (132 mg) was dissolved in DMF (5 mL) and sequentially Injections (0.4 mL each) were purified by preparative HPLC. The product (-) - (1'S) -53a (95.3 mg, 107 μmol, 88%) was obtained as a light yellow solid. HPLC (preparative): Pillar: Kromasil 100 C18, 250 × 20 mm, 7 μm Guard column: Kromasil 100 C18, 50 × 20 mm, 5 μm Gradient: Time [min] H ₂ O / MeOH 0 30/70 0-15 70/30 → 0/100 15-22 0/100 22-23 0/100 → 70/30 23-30 30/70 River: 16 mL min ^-1 λ: 254 nm t _R : 13.9 min [Α] 20 / D = -43.6 ° (c = 0.257, DMSO).
UV (CH ₃ CN): λ _max (Ig ε) = 209.0 nm (4,447), 254.0 (4,702) 262.5 (4,642) 304.0 (4,120) 316.0 (4,200).
IR (KBr): ṽ (cm ^-1 ) = 3406, 2927, 1655, 1580, 1469, 1403, 1313, 1263, 1131, 1076, 856, 760, 664.
¹ H-NMR (600 MHz, DMSO-d ₆ ): δ = 1.99 (m _c , 2H, 3-H ₂ ), 2.60-2.65 (m, 2H, 2-H _a , 4-H _a ), 2.69- 2.74 (m, 2H, 2H _b, 4-H _b), 3:46 to 3:49 (m, 2H, 2x3 '' - H), 3.52-3.64 (m, 6H, 2x5 '' - H, 2x6 '' - H ₂ ), 3.75-3.79 (m, 4H, 2x2 "-H, 2x4" -H), 3.85-3.88 (m, 2H, 2x10'-H _a ), 4.02 (dd, J = 10.8, 2.4 Hz , 2H, 2x10'-H _b), 4:22 to 4:24 (m, 4H, H-2x1', 2x2'-H _a), 4:37 to 4:40 (m, 2H, 2x2'-H _b), 4:51 (d, J = 4.6 Hz, 2H, 2xOH), 4:54 (m _c, 2H, 2xOH), 4.82 (d, J = 5.5 Hz, 2H, 2xOH), 4.93 (d, J = 7.1 Hz, 2H, 2x1 '' - H) , 5.27 (d, J = 4.5 Hz, 2H, 2xOH), 7.39 (t, J = 7.7 Hz, 2H, 2x7'-H), 7.54 (t, J = 7.5 Hz, 2H, 2x8'-H), 7.85 (d, J = 8.6 Hz, 2H, 2x9'-H), 8.30 (d, J = 8.6 Hz, 2H, 2x6'-H), 8.23 (s, 2H, 2x4'-H).
¹³ C-NMR (125 MHz, DMSO-d ₆ ): δ = 19.2 (C-3), 34.5 (C-2, C-4), 40.7 (2 × C-1 '), 47.6 (2 × C-10'), 52.6 (2xC-2 '), 59.7 (2xC-6''), 67.6 (2xC-4''), 70.4 (2xC-2''), 73.1 (2xC-3''), 75.2 (2xC-5''), 101.2 (2xC-4'), 101.9 (2xC-1 ''), 116.6 (2xC-5a '), 122.4 (2xC-9', 2xC-9b '), 123.2 (2xC-6', 2xC- 7 '), 127.2 (2xC-8'), 129.4 (2xC-9a '), 141.7 (2xC-3a'), 153.7 (2xC-5 '), 170.6 (2xC = 0).
MS (ESI): m / z (%) = 909.2 (47) [M + Na] ⁺ , 885.3 (100) [MH] ^- . C ₄₃ H ₄₈ Cl ₂ N ₂ O ₁₄ (887.75). ber .: 909.2375 Found: 909.2377, [M + Na] ⁺ (ESI-HRMS).

Example 5

1,6-bis [(1S) -1-chloromethyl-5-hydroxy-1,2-dihydro-3H-benz [e] indol-3-yl] hexane-1,6-dione]} ((1'S) - 55b, n = 4)

The phenol (-) - (1S) -69 (58.0 mg, 174 μmol, 2.0 eq.) Was dissolved in 4 M HCl / ethyl acetate (4.5 mL) and after stirring at room temperature for 1 h the solvent was removed in vacuo. The residue was dried in vacuo for 1 h, dissolved in DMF (5 mL), the solution was cooled to 0 ° C. and admixed with adipic acid chloride (66b) (12.9 μL, 15.9 mg, 86.9 μmol, 1.0 eq.) And pyridine (28.0 μL, 27.5 mg, 348 μmol, 4.0 eq.). After stirring for 20 h at room temperature, the reaction mixture was diluted with DMF (5 mL) and purified in successive injections (0.4 mL each) by preparative HPLC. 3 mg of a mixture of the product (1'S) -55b (<5.19 μmol, <6%) and not clearly identified by-products were obtained as a white solid. HPLC (preparative): Pillar: Kromasil 100 C18, 250 × 20 mm, 7 μm Guard column: - Gradient: Time [min] H ₂ O / MeOH 0 30/70 0-6.0 30/70 → 0/100 6.5-8.5 0/100 8.5-9.5 0/100 → 30/70 9.5-15 30/70 River: 16 mL min ^-1 λ: 254 nm t _R : 9.4 min ¹ H-NMR (600 MHz, DMSO-d ₆ ): δ = 1.74 (m _c , 4H, 3-H ₂ , 4-H ₂ ), 2.57 (m _c , 4H, 2-H ₂ , 5-H ₂ ), 3.79 (dd, J = 10.9, 7.9 Hz, 2H, 2x10'-H _a ), 3.98 (dd, J = 11.1, 3.1 Hz, 2H, 2x10'-H _b ), 4.13-4.19 (m, 4H, 2x1'-H, 2x2'-H _a ), 4.30-4.36 (m, 2H, 2x2'-H _b ), 7.31 (t, J = 7.6 Hz, 2H, 2x7'-H), 7.48 (t, J = 7.6Hz, 2H, 2x8'-H), 7.76 (d, J = 8.3Hz, 2H, 2x9'-H), 7.99 (s, 2H, 2x4'-H), 8.07 (d, J = 8.4Hz , 2H, 2x6'-H), 10.32 (s, 2H, 2xOH).
¹³ C-NMR (125 MHz, DMSO-d ₆ ): δ = 23.8 (C-3, C-4), 35.0 (C-2, C-5), 40.7 (2 × C-1 '), 47.7 (2 × C) 10 '), 52.7 (2xC-2'), 99.8 (2xC-4 '), 113.6 (2xC-5a'), 121.5 (2xC-9b '), 122.4 (2xC-9'), 122.5 (2xC-7 ' ), 123.0 (2xC-6 '), 127.0 (2xC-8'), 129.8 (2xC-9a '), 141.9 (2xC-3a'), 154.1 (2xC-5 '), 170.6 (2xC = O).
MS (ESI): m / z (%) = 575.2 (9) [MH] ^- , 539.2 (26) [M-2H-Cl] ^- . C ₃₂ H ₃₀ Cl ₂ N ₂ O ₄ (577.50). ber .: 575.1510 Found: 575.1520, [MH] ^- (ESI-HRMS).

Example 6

(-) - {1,6-bis [(1S) -1-chloromethyl-5- (β-D-galactopyranosyl) -1,2-dihydro-3H-benz [e] indol-3-yl] hexan-1 , 6-dione]} ((-) - (1'S) -53b, n = 4)

The acetyl protected galactoside (1'S) -71b (125 mg, 121 μmol, 1.0 eq.) Was suspended in absolute methanol (35 mL) and washed at 0 ° C with NaOMe in methanol (75 μL of a 5.4 M solution, 404 μmol, 4.0 eq .). After warming to room temperature and stirring for 2 h at this temperature the suspension with water (3.0 mL) was diluted and so long as acidic ion exchanger (Amberlite IR-120 ^®) were added until the mixture was neutral (pH = 7). The mixture was separated via transfer cannula from the ion exchanger, the same washed with DMF (2 × 6 mL), the organic phases combined and the solvent removed in vacuo. The resulting residue (123 mg) was dissolved in DMF (5 mL) and purified in successive injections (0.4 mL each) by preparative HPLC. The product (-) - (1'S) -53b (76.2 mg, 84.5 μmol, 84%) was obtained as a light yellow solid. HPLC (preparative): Pillar: Kromasil 100 C18, 250 × 20 mm, 7 μm Guard column: Kromasil 100 C18, 50 × 20 mm, 5 μm Gradient: Time [min] H ₂ O / MeOH 0 30/70 0-15 70/30 → 0/100 15-22 0/100 22-23 0/100 → 70/30 23-30 30/70 River: 16 mL min ^-1 λ: 254 nm t _R : 14.4 min [Α] 20 / D = -45.0 ° (c = 0.249, DMSO).
UV (CH ₃ CN): λ _max (Ig ε) = 208.0 nm (4,363), 254.5 (4,623) 262.5 (4,630) 304.0 (3,998) 315.0 (4,055).
IR (KBr): ṽ (cm ^-1 ) = 3406, 2927, 1656, 1580, 1469, 1402, 1316, 1075, 760, 665.
¹ H-NMR (600 MHz, DMSO-d ₆ ): δ = 1.75 (m _c , 4H, 3-H ₂ , 4-H ₂ ), 2.60 (m _c , 4H, 2-H ₂ , 5-H ₂ , 3.48-3.54 (m, 4H, 2x3 "-H, 2x6" -H _a ), 3.56-3.64 (m, 4H, 2x5 "-H, 2x6" -H _b ), 3.75-3.80 ( m, 4H, 2x2 '' - H, 2x4 '' - H), 3.86 (m _c, 2H, 2x10'-H _a), 4:02 (dd, J = 11.0, 2.7 Hz, 2H, H-2x10' _b) , 4:22 to 4:23 (, 2x2'-H m, 4H, 2x1'-H _a), 4:38 to 4:41 (m, 2H, 2x2'-H _b), and 4.50 (d, J = 4.6 Hz, 2H, 2xOH), 4:53 (m _c, 2H, 2xOH), 4.82 (d, J = 5.6 Hz, 2H, 2xOH), 4.93 (d, J = 7.1 Hz, 2H, 2x1 '' - H), 5.26 (d, J = 5.1 Hz , 2H, 2xOH), 7.38 (t, J = 7.7Hz, 2H, 2x7'-H), 7.54 (t, J = 7.6Hz, 2H, 2x8'-H), 7.85 (d, J = 8.4Hz, 2H , 2x9'-H), 8.29-8.30 (m, 4H, 2x4'-H, 2x6'-H).
¹³ C-NMR (125 MHz, DMSO-d ₆ ): δ = 23.6 (C-3, C-4), 35.0 (C-2, C-5), 40.7 (2 × C-1 '), 47.6 (2 × C) 10 '), 52.6 (2xC-2'), 59.6 (2xC-6 ''), 67.6 (2xC-4 ''), 70.4 (2xC-2 ''), 73.1 (2xC-3 ''), 75.2 ( 2xC-5 ''), 101.2 (2xC-4 '), 101.9 (2xC-1''), 116.5 (2xC-5a'), 122.4 (2xC-9 ', 2xC-9b'), 123.1 (2xC-6 ', 2xC-7'), 127.2 (2xC-8 '), 129.4 (2xC-9a'), 141.7 (2xC-3a '), 153.6 (2xC-5'), 170.7 (2xC = 0).
MS (ESI): m / z (%) = 923.3 (67) [M + Na] ⁺ , 899.3 (100) [MH] ^- . C ₄₄ H ₅₀ Cl ₂ N ₂ O ₁₄ (901.78). ber .: 923.2531 Found: 923.2530, [M + Na] ⁺ (ESI-HRMS).

Example 7

(-) - {1,7-bis [(1S) -1-chloromethyl-5-hydroxy-1,2-dihydro-3H-benz [e] indol-3-yl] heptane-1,7-dione]} ((-) - (1'S) -55c, n = 5)

The phenol (-) - (1S) -69 (62.0 mg, 186 μmol, 2.0 eq.) Was dissolved in 4 M HCl / ethyl acetate (4.5 mL) and, after stirring at room temperature for 1 h, the solvent was removed in vacuo. The residue was dried in vacuo for 1 h, dissolved in DMF (5 mL), the solution was cooled to 0 ° C. and treated with pimelic acid chloride (66c) (15.5 μL, 18.3 mg, 92.9 μmol, 1.0 eq.) And pyridine (30.0 μL, 29.4 mg, 371 μmol, 4.0 eq.). After stirring for 20 h at room temperature, the reaction mixture was diluted with DMF (5 mL) and purified in successive injections (0.4 mL each) by preparative HPLC. The product (-) - (1'S) -55c (35.9 mg, 60.7 μmol, 65%) was obtained as a white solid. HPLC (preparative): Pillar: Kromasil 100 C18, 250 × 20 mm, 7 μm Guard column: - Gradient: Time [min] H ₂ O / MeOH 0 30/70 0-6.0 30/70 → 0/100 6.0-8.5 0/100 8.5-9.5 0/100 → 30/70 9.5-15 30/70 River: 16 mL min ^-1 λ: 254 nm t _R : 8.9 min [Α] 20 / D = -33.6 ° (c = 0.268, DMSO).
UV (CH ₃ CN): λ _max (Ig ε) = 256.0 nm (4.531), 263.5 (4.514), 318.0 (4.006). ¹
IR (KBr): ṽ (cm ^-1 ) = 3193, 2925, 1636, 1582, 1520, 1474, 1424, 1391, 1247, 1129, 1062, 851, 774, 718.
¹ H-NMR (600 MHz, DMSO-d ₆ ): δ = 1.46-1.51 (m, 2H, 4-H ₂ ), 1.70 (m _c , 4H, 3-H ₂ , 5-H ₂ ), 2.53 ( m _c , 4H, 2-H ₂ , 6-H ₂ ), 3.78 (dd, J = 10.9, 8.1 Hz, 2H, 2x10'-H _a ), 3.98 (dd, J = 11.1, 3.0 Hz, 2H, 2x10 '-H _b ), 4.12-4.18 (m, 4H, 2x1'-H 2x2'-H _a ), 4.34 (m _c , 2H, 2x2'-H _b ), 7.31 (m _c , 2H, 2x7'-H ), 7:48 (m _c, 2H, 2x8'-H), 7.77 (d, J = 8.4 Hz, 2H, 2x9'-H), 8:00 (s, 2H, 2x4'-H), 8:08 (d, J = 8.4 Hz, 2H, 2x6'-H), 10.28 (s, 2H, 2xOH).
¹³ C-NMR (125 MHz, DMSO-d ₆ ): δ = 24.0 (C-3, C-5), 28.3 (C-4), 35.0 (C-2, C-6), 40.7 (2xC-1 '), 47.6 (2xC-10'), 52.6 (2xC-2 '), 99.7 (2xC-4'), 113.5 (2xC-5a '), 121.4 (2xC-9b'), 122.3 (2xC-9 ') , 122.4 (2xC-7 '), 122.9 (2xC-6'), 126.9 (2xC-8 '), 129.8 (2xC-9a'), 141.9 (2xC-3a '), 154.0 (2xC-5'), 170.6 (2xC = O).
MS (ESI): m / z (%) = 1181.5 (6) [2M-H] ^- , 589.2 (89) [MH] ^- , 553.2 (86) [M-2H-Cl] ^- . C ₃₃ H ₃₂ Cl ₂ N ₂ O ₄ (591.52). calculated: 613.1631 Found: 613.1634, [M + Na] ⁺ (ESI-HRMS).

Example 8

(-) - {1,7-bis [(1S) -1-chloromethyl-5 - ([β-D-galactopyranosyl) -1,2-dihydro-3H-benz [e] indol-3-yl] heptane 1,7-dione]} ((-) - (1'S) -53c, n = 5),

The acetyl protected galactoside (1'S) -71c (152 mg, 121 μmol, 1.0 eq.) Was suspended in absolute methanol (35 mL) and washed at 0 ° C with NaOMe in methanol (90 μL of a 5.4 M solution, 486 μmol, 4.0 eq .). After warming to room temperature and stirring for 45 minutes at this temperature, the substrate had completely dissolved. Stirring was continued for a further 1.5 h at room temperature, the reaction solution followed by water (3.0 mL) and as long as acidic ion exchanger added (Amberlite IR-120 ^®), until the solution was neutral (pH = 7). The mixture was separated via transfer cannula from the ion exchanger, the same washed with methanol (2 × 6 mL), the organic phases combined and the solvent removed in vacuo. The resulting residue (142 mg) was dissolved in DMF (5 mL) and sequentially Injections (0.3 mL each) are purified by preparative HPLC. The product (-) - (1'S) -53c (98.6 mg, 108 μmol, 89%) was obtained as a pale yellow solid. HPLC (preparative): Pillar: Kromasil 100 C18, 250 × 20 mm, 7 μm Guard column: Kromasil 100 C18, 50 × 20 mm, 5 μm Gradient: Time [min] H ₂ O / MeOH 0 30/70 0-15 70/30 → 0/100 15-22 0/100 22-23 0/100 → 70/30 23-30 30/70 River: 16 mL min ^-1 λ: 254 nm t _R : 15.0 min [Α] 20 / D = -42.9 ° (c = 0.252, DMSO).
UV (CH ₃ CN): λ _max (Ig ε) = 208.0 nm (4,521), 254.5 (4,798) 262.0 (4,801) 303.5 (4,159) 315.0 (4,219).
IR (KBr): ṽ (cm ^-1 ) = 3406, 2929, 1656, 1580, 1469, 1402, 1313, 1131, 1075, 760, 664.
¹ H-NMR (600 MHz, DMSO-d ₆ ): δ = 1.49 (m _c , 2H, 4-H ₂ ), 1.71 (m _c , 4H, 3-H ₂ , 5-H ₂ ), 2.55 (m _c , 4H, 2-H ₂ , 6 -H ₂ ), 3.48-3.53 (m, 4H, 2x3 "-H, 2x6" -H _a ), 3.58-3.64 (m, 4H, 2x5 "-H , 2x6 '' - H _b), 3.75-3.81 (m, 4H, 2x2 '' - H, 2x4 '' - H), 3.86 (m _c, 2H, 2x10'-H _a), 4:02 (dd, J = 11.0, 2.7 Hz, 2H, 2x10'-H _b), 4:21 to 4:22 (m, 4H, H-2x1', 2x2'-H _a), 4:36 to 4:40 (m, 2H, 2x2'-H _b), 4:51 (d, J = 4.6 Hz, 2H, 2xOH), 4:54 (m _c, 2H, 2xOH), 4.83 (d, J = 5.5 Hz, 2H, 2xOH), 4.94 (d, J = 7.3 Hz, 2H, 2x1 ''-H), 5.27 (d, J = 5.0Hz, 2H, 2xOH), 7.38 (t, J = 7.7Hz, 2H, 2x7'-H), 7.54 (t, J = 7.5Hz, 2H, 2x8'- H), 7.85 (d, J = 8.4 Hz, 2H, 2x9'-H), 8.29-8.30 (m, 4H, 2x4'-H, 2x6'-H).
¹³ C-NMR (125 MHz, DMSO-d ₆ ): δ = 23.9 (C-3, C-5), 28.3 (C-4), 35.1 (C-2, C-6), 40.7 (2xC-1 '), 47.6 (2xC-10'), 52.6 (2xC-2 '), 59.6 (2xC-6''), 67.6 (2xC-4''), 70.4 (2xC-2''), 73.1 (2xC-2') 3 ''), 75.2 (2xC-5 ''), 101.2 (2xC-4 '), 101.9 (2xC-1''), 116.6 (2xC-5a'), 122.4 (2xC-9 ', 2xC-9b' ), 123.2 (2xC-6 ', 2xC-7'), 127.2 (2xC-8 '), 129.4 (2xC-9a'), 141.7 (2xC-3a '), 153.6 (2xC-5'), 170.8 (2xC = O).
MS (ESI): m / z (%) = 937.3 (69) [M + Na] ⁺ , 913.3 (100) [MH] ^- . C ₄₅ H ₅₂ Cl ₂ N ₂ O ₁₄ (915.81). ber .: 937.2688 Found: 937.2691, [M + Na] ⁺ (ESI-HRMS).

Example 9

(+) - {1,5-bis [(1S, 10R) -1- (10-Chloroethyl) -5-hydroxy-1,2-dihydro-3H-benz [e] indol-3-yl] pentan-1 , 5-dione]} ((+) - (1'S, 10'R) -56a, n = 3)

wurde in 4 M HCl/Essigester (4.5 mL) gelöst und nach 1 h Rühren bei Raumtemperatur das Lösungsmittel im Vakuum entfernt. Der Rückstand wurde 1 h im Vakuum getrocknet, in DMF (5 mL) gelöst, die Lösung auf 0°C gekühlt und mit Glutarsäurechlorid (66a) (11.7 μL, 15.1 mg, 89.3 μmol, 1.0 Äq.) sowie Pyridin (28.8 μL, 28.2 mg, 357 μmol, 4.0 Äq.) versetzt. Nach 20 h Rühren bei Raumtemperatur wurde das Reaktionsgemisch mit DMF (5 mL) verdünnt und in aufeinanderfolgenden Injektionen (jeweils 0.4 mL) mittels präparativer HPLC gereinigt. Das Produkt (+)-(1'S,10'R)-56a (42.1 mg, 71.2 μmol, 80%) wurde als weißer Feststoff erhalten. HPLC (präparativ): Säule: Kromasil 100 C18, 250 × 20 mm, 7 –m Vorsäule: Kromasil 100 C18, 50 × 20 mm, 5 μm Gradient: Zeit [min] H₂O/MeOH 0 30/70 0–6.0 30/70 → 0/100 6.0–8.5 0/100 8.5–9.5 0/100 → 30/70 9.5–15 30/70 Fluss: 16 mL min^–1 λ: 254 nm t_R: 8.7 min [α] 20 / D = +23.2° (c = 0.25, DMSO).
UV (CH₃CN): λ_max (lg ε) = 206.5 nm (4.576), 222.5 (4.410), 257.0 (4.948), 265.0 (4.977), 320.5 (4.355).
IR (KBr): ṽ (cm^–1) = 3199, 1632, 1684, 1522, 1453, 1415, 1394, 1263, 1158, 1134, 1074, 1024, 859, 757.
¹H-NMR (600 MHz, DMSO-d₆): δ = 1.64 (d, J = 6.6 Hz, 6H, 2x10'-CH₃), 1.99 (m_c, 2H, 3-H₂), 2.58-2.63 (m, 2H, 2-H_a, 4-H_a), 2.70-2.75 (m, 2H, 2-H_b, 4-H_b), 4.11-4.12 (m, 2H, 2x1'-H), 4.24-4.29 (m, 4H, 2x2'-H₂), 4.78 (m_c, 2H, 2x10'-H), 7.31 (t, J = 7.6 Hz, 2H, 2x7'-H), 7.47 (t, J = 7.5 Hz, 2H, 2x8'-H), 7.82 (d, J = 8.4 Hz, 2H, 2x9'-H), 8.01 (s, 2H, 2x4'-H), 8.10 (d, J = 8.4 Hz, 2H, 2x6'-H), 10.25 (s, 2H, 2xOH).
¹³C-NMR (125 MHz, DMSO-d₆): δ = 19.6 (C-3), 23.2 (2xC-11'), 34.5 (C-2, C-4), 45.2 (2xC-1'), 49.6 (2xC-2'), 61.5 (2xC-10'), 99.7 (2xC-4'), 114.7 (2xC-5a'), 121.5 (2xC-9b'), 122.3 (2xC-7'), 122.5 (2xC-9'), 122.9 (2xC-6'), 126.7 (2xC-8'), 129.7 (2xC-9a'), 141.8 (2xC-3a'), 153.7 (2xC-5'), 170.1 (2xC=O).
MS (ESI): m/z (%) = 1205.3 (100) [2M+Na]⁺, 907.2 (23) [3M+2Na]²⁺, 613.1 (43) [M+Na]⁺, 589.2 (100) [M-H]^–. C₃₃H₃₂Cl₂N₂O₄ (591.52). ber.: 589.1666 gef.: 589.1694, [M-H]^– (ESI-HRMS). The phenol (+) - (1S, 10R) -109 (62.1 mg, 179 μmol, 2.0 eq.)

was dissolved in 4 M HCl / ethyl acetate (4.5 mL) and after stirring at room temperature for 1 h the solvent was removed in vacuo. The residue was dried in vacuo for 1 h, dissolved in DMF (5 mL), the solution was cooled to 0 ° C. and treated with glutaric acid chloride (66a) (11.7 μL, 15.1 mg, 89.3 μmol, 1.0 eq.) And pyridine (28.8 μL, 28.2 mg, 357 μmol, 4.0 eq.). After stirring for 20 h at room temperature, the reaction mixture was diluted with DMF (5 mL) and purified in successive injections (0.4 mL each) by preparative HPLC. The product (+) - (1'S, 10'R) -56a (42.1 mg, 71.2 μmol, 80%) was obtained as a white solid. HPLC (preparative): Pillar: Kromasil 100 C18, 250 × 20 mm, 7 -m Guard column: Kromasil 100 C18, 50 × 20 mm, 5 μm Gradient: Time [min] H ₂ O / MeOH 0 30/70 0-6.0 30/70 → 0/100 6.0-8.5 0/100 8.5-9.5 0/100 → 30/70 9.5-15 30/70 River: 16 mL min ^-1 λ: 254 nm t _R : 8.7 min [Α] 20 / D = + 23.2 ° (c = 0.25, DMSO).
UV (CH ₃ CN): λ _max (Ig ε) = 206.5 nm (4,576), 222.5 (4,410) 257.0 (4,948) 265.0 (4,977) 320.5 (4,355).
IR (KBr): ṽ (cm ^-1 ) = 3199, 1632, 1684, 1522, 1453, 1415, 1394, 1263, 1158, 1134, 1074, 1024, 859, 757.
¹ H-NMR (600 MHz, DMSO-d ₆ ): δ = 1.64 (d, J = 6.6 Hz, 6H, 2x10'-CH ₃ ), 1.99 (m _c , 2H, 3-H ₂ ), 2.58-2.63 (m, 2H, 2H _a, 4-H _a), 2.70-2.75 (m, 2H, 2H _b, 4-H _b), 4:11 to 4:12 (m, 2H, 2x1'-H), 4.24 -4.29 (m, 4H, 2x2'-H ₂ ), 4.78 (m _c , 2H, 2x10'-H), 7.31 (t, J = 7.6 Hz, 2H, 2x7'-H), 7.47 (t, J = 7.5 Hz, 2H, 2x8'-H), 7.82 (d, J = 8.4 Hz, 2H, 2x9'-H), 8.01 (s, 2H, 2x4'-H), 8.10 (d, J = 8.4 Hz, 2H , 2x6'-H), 10.25 (s, 2H, 2xOH).
¹³ C-NMR (125 MHz, DMSO-d ₆ ): δ = 19.6 (C-3), 23.2 (2 × C-11 '), 34.5 (C-2, C-4), 45.2 (2 × C-1'), 49.6 (2xC-2 '), 61.5 (2xC-10'), 99.7 (2xC-4 '), 114.7 (2xC-5a'), 121.5 (2xC-9b '), 122.3 (2xC-7'), 122.5 ( 2xC-9 '), 122.9 (2xC-6'), 126.7 (2xC-8 '), 129.7 (2xC-9a'), 141.8 (2xC-3a '), 153.7 (2xC-5'), 170.1 (2xC = O).
MS (ESI): m / z (%) = 1205.3 (100) [2M + Na] ⁺ , 907.2 (23) [3M + 2Na] ²⁺ , 613.1 (43) [M + Na] ⁺ , 589.2 (100) [MH] ^- . C ₃₃ H ₃₂ Cl ₂ N ₂ O ₄ (591.52). ber .: 589.1666 Found: 589.1694, [MH] ^- (ESI-HRMS).

Example 10

(-) - {1,5-bis [(1S, 10R) -1- (10-chloroethyl) -5- (β-D-galactopyranosyl) -1,2-dihydro-3H-benz [e] indol-3 -yl] pentane-1,5-dione]} ((-) - (1'S, 10'R) -54a, n = 3)

A mixture (96.3 mg) of the acetyl protected galactoside (1'S, 10'R) -72a and an ambiguously identified by-product was suspended in absolute methanol (30 mL) and NaOMe in methanol (57 μL of a 5.4 M solution , 308 μmol). After warming to room temperature and stirring for 30 min at this temperature, the substance mixture had completely dissolved. Stirring was continued for a further 1.5 h at room temperature, the reaction solution followed by water (3.0 mL) and as long as acidic ion exchanger added (Amberlite IR-120 ^®), until the solution was neutral (pH = 7). The mixture was separated via transfer cannula from the ion exchanger, the same washed with methanol (2 × 6 mL), the organic phases combined and the solvent removed in vacuo. The residue obtained (68.2 mg) was dissolved in DMF (5 mL) and purified in successive injections (0.4 mL each) by preparative HPLC. The product (-) - (1'S, 10'R) -54a (44.7 mg, 48.8 μmol, 30% over 3 stages and twice purification by preparative HPLC) was obtained as a light yellow solid. HPLC (preparative): Pillar: Kromasil 100 C18, 250 × 20 mm, 7 -m Guard column: Kromasil 100 C18, 50 × 20 mm, 5 μm Gradient: Time [min] H ₂ O / MeOH 0 30/70 0-15 70/30 → 0/100 15-22 0/100 22-23 0/100 → 70/30 23-30 30/70 River: 16 mL min ^-1 λ: 254 nm t _R : 12.4 min [Α] 20 / D = -7.2 ° (c = 0.25, DMSO).
UV (CH ₃ CN): λ _max (Ig ε) = 209.0 nm (4,221), 254.5 (4,531), 263.0 (4,489), 306.5 (3,945), 316.5 (4,021).
IR (KBr): ṽ (cm ^-1 ) = 3406, 2892, 1627, 1581, 1517, 1470, 1402, 1209, 1135, 1075, 854, 760, 618.
¹ H-NMR (600 MHz, DMSO-d ₆ ): δ = 1.66 (d, J = 6.3 Hz, 6H, 2x10'-CH ₃ ), 2.00 (m _c , 2H, 3-H ₂ ), 2.60-2.68 (m, 2H, 2H _a, 4-H _a), 2.69-2.76 (m, 2H, 2H _b, 4-H _b), 3:45 to 3:50 (m, 2H, 2x3 '' - H), 3.52-3.66 (m, 6H, 2x5 "-H, 2x6" -H ₂ ), 3.75-3.82 (m, 4H, 2x2 "-H, 2x4" -H), 4.18-4.22 (m, 2H , 2x1'-H), 4.26-4.36 (m, 4H, 2x2'-H ₂ ), 4.52 (d, J = 4.6 Hz, 2H, 2xOH), 4.55 (m _C , 2H, 2xOH), 4.80-4.87 ( m, 4H, 2x10'-H, 2xOH), 4.92 (d, J = 7.5Hz, 2H, 2x1 "-H), 5.27 (d, J = 4.9Hz, 2H, 2xOH), 7.38 (t, J = 7.5Hz, 2H, 2x7'-H), 7.53 (t, J = 7.5Hz, 2H, 2x8'-H), 7.90 (d, J = 8.4Hz, 2H, 2x9'-H), 8.31-8.32 (m , 4H, 2x4'-H, 2x6'-H).
¹³ C-NMR (125 MHz, DMSO-d ₆ ): δ = 19.4 (C-3), 23.1 (2 × C-11 '), 34.5 (C-2, C-4), 45.3 (2 × C-1'), 49.5 (2xC-2 '), 59.7 (2xC-6''), 61.3 (2xC-10'), 67.6 (2xC-4 ''), 70.5 (2xC-2 ''), 73.2 (2xC-3 '' ), 75.3 (2xC-5 ''), 101.3 (2xC-4 '), 102.1 (2xC-1 ''), 117.8 (2xC-5a '), 122.6 (2 signals) (2xC-9', 2xC-9b '), 123.2 (2xC-7'), 123.3 (2xC-6 '), 127.1 (2xC-8 '), 129.5 (2xC-9a'), 141.7 (2xC-3a '), 153.6 (2xC-5'), 170.5 (2xC = O).
MS (ESI): m / z (%) = 1394.4 (8) [3M + 2Na] ²⁺ , 937.3 (100) [M + Na] ⁺ . C ₄₅ H ₅₂ Cl ₂ N ₂ O ₁₄ (915.81). ber .: 937.2685 Found: 937.2688, [M + Na] ⁺ (ESI-HRMS).

Example 11

(+) - {1,6-bis [(1S, 10R) -1- (10-Chloroethyl) -5-hydroxy-1,2-dihydro-3H-benz [e] endol-3-yl] hexan-1 , 6-dione]} ((+) - (1'S, 10'R) -56b, n = 4)

The phenol (+) - (1S, 10R) -109 (59.7 mg, 172 μmol, 2.0 eq.) Was dissolved in 4 M HCl / ethyl acetate (4.5 mL) and, after stirring at room temperature for 1 h, the solvent was removed in vacuo. The residue was dried in vacuo for 1 h, dissolved in DMF (5 mL), the solution was cooled to 0 ° C. and admixed with adipic acid chloride (66b) (12.5 μL, 15.7 mg, 85.8 μmol, 1.0 eq.) And pyridine (27.7 μL, 27.2 mg, 343 μmol, 4.0 eq.). After stirring for 20 h at room temperature, the reaction mixture was diluted with DMF (5 mL) and purified in successive injections (0.4 mL each) by preparative HPLC. The product (+) - (1'S, 10'R) -56b (39.4 mg, 65.1 μmol, 76%) was obtained as a white solid. HPLC (preparative): Pillar: Kromasil 100 C18, 250 × 20 mm, 7 -m Guard column: - Gradient: Time [min] H ₂ O / MeOH 0 30/70 0-6.0 30/70 → 0/100 6.0-8.5 0/100 8.5-9.5 0/100 → 30/70 9.5-15 30/70 River: 16 mL min ^-1 λ: 254 nm t _R : 8.8 min [Α] 20 / D = + 8.2 ° (c = 0.269, DMSO).
UV (CH ₃ CN): λ _max (Ig ε) = 196.5 nm (4,295), 256.5 (4,171), 264.5 (4,177), 320.0 (3,541).
IR (KBr): ṽ (cm ^-1 ) = 3210, 2924, 1665, 1633, 1582, 1522, 1394, 1253, 1135, 861, 750.
¹ H-NMR (600 MHz, DMSO-d ₆ ): δ = 1.63 (d, J = 6.6 Hz, 6H, 2x10'-CH ₃ ), 1.76 (m _c , 4H, 3-H ₂ , 4-H ₂ ), 2:53 to 2:59 (m, 2H, 2H _a, 5-H _a), 2.60-2.66 (m, 2H, 2H _b, 5-H _b), 4.10 (m _c, 2H, 2x1'- H), 4.26-4.27 (m, 4H, 2x2'-H ₂ ), 4.77 (m _c , 2H, 2x10'-H), 7.30 (t, J = 7.6 Hz, 2H, 2x7'-H), 7.46 ( t, J = 7.5 Hz, 2H, 2x8'-H), 7.81 (d, J = 8.6 Hz, 2H, 2x9'-H), 7.99 (s, 2H, 2x4'-H), 8.09 (d, J = 8.4 Hz, 2H, 2x6'-H), 10.22 (s, 2H, 2xOH).
¹³ C-NMR (125 MHz, DMSO-d ₆ ): δ = 23.2 (2 × C-11 '), 23.8 (C-3, C-4), 35.0 (C-2, C-5), 45.2 (2 × C) 1 '), 49.5 (2xC-2'), 61.5 (2xC-10 '), 99.7 (2xC-4'), 114.6 (2xC-5a '), 121.5 (2xC-9b'), 122.2 (2xC-7 ' ), 122.5 (2xC-9 '), 122.9 (2xC-6'), 126.6 (2xC-8 '), 129.7 (2xC-9a'), 141.8 (2xC-3a '), 153.7 (2xC-5'), 170.2 (2xC = O).
MS (ESI): m / z (%) = 627.2 (26) [M + Na] ⁺ , 603.2 (80) [MH] ^- , 567.3 (100) [M-2H-Cl] ^- . C ₃₄ H ₃₄ Cl ₂ N ₂ O ₄ (605.55). ber .: 603.1823 Found: 603.1837, [MH] ^- (ESI-HRMS).

Example 12

(-) - {1,6-bis [(1S, 10R) -1- (10-chloroethyl) -5- (β-D-galactopyranosyl) -1,2-dihydro-3H-benz [e] indol-3 -yl] hexane-1,6-dione]} ((-) - (1'S, 10'R) -54b, n = 4)

A mixture (97.3 mg) of the acetyl protected galactoside (1'S, 10'R) -72b and an ambiguously identified by-product was suspended in absolute methanol (30 mL) and NaOMe in methanol (57 μL of a 5.4 M solution) at 0 ° C , 307 μmol). After warming to room temperature and stirring for 30 min at this temperature, the substance mixture had completely dissolved. Stirring was continued for a further 1.5 h at room temperature, the reaction solution followed by water (3.0 mL) and as long as acidic ion exchanger added (Amberlite IR-120 ^®), until the solution was neutral (pH = 7). The mixture was separated via transfer cannula from the ion exchanger, the same washed with methanol (2 × 6 mL), the organic phases combined and the solvent removed in vacuo. The resulting residue (75.9 mg) was dissolved in DMF (5 mL) and purified in successive injections (0.3 mL) by preparative HPLC. The product (-) - (1'S, 10'R) -54b (43.9 mg, 47.2 μmol, 27% over 3 steps) was obtained as a light yellow solid. HPLC (preparative): Pillar: Kromasil 100 C18, 250 × 20 mm, 7 μm Guard column: Kromasil 100 C18, 50 × 20 mm, 5 μm Gradient: Time [min] H ₂ O / MeOH 0 30/70 0-15 70/30 → 0/100 15-22 0/100 22-23 0/100 → 70/30 23-30 30/70 River: 16 mL min ^-1 λ: 254 nm t _R : 13.4 min [Α] 20 / D = -13.0 ° (c = 0.269, DMSO).
UV (CH ₃ CN): λ _max (Ig ε) = 199.0 nm (3,990), 255.0 (3,268), 262.5 (3,255), 304.0 (2,634), 313.5 (2,649).
IR (KBr): ṽ (cm ^-1 ) = 3406, 2925, 1651, 1579, 1469, 1414, 1210, 1139, 1076, 857, 761.
¹ H-NMR (600 MHz, DMSO-d ₆ ): δ = 1.65 (d, J = 6.7 Hz, 6H, 2x10'-CH ₃ ), 1.77 (m _C , 4H, 3-H ₂ , 4-H ₂ ), 2.63 (m _c , 4H, 2-H ₂ , 5-H ₂ ), 3.47-3.55 (m, 4H, 2x3 "-H, 2x6" -H _a ), 3.57-3.64 (m, 4H, 2x5 '' - H, 2x6 '' - H _b), 3.74-3.81 (m, 4H, 2x2 '' - H, 2x4 '' - H), 4:16 to 4:21 (m, 2H, 2x1'-H), 4.28 -4.34 (m, 4H, 2x2'-H ₂ ), 4.51 (d, J = 4.0Hz, 2H, 2xOH), 4.54 (m _C , 2H, 2xOH), 4.80-4.85 (m, 4H, 2x10'-H , 2xOH), 4.92 (d, J = 7.5 Hz, 2H, 2x1 "-H), 5.26 (s _br , 2H, 2xOH), 7.37 (t, J = 7.6 Hz, 2H, 2x7'-H), 7.52 (t, J = 7.5 Hz, 2H, 2x8'-H), 7.88 (d, J = 8.4 Hz, 2H, 2x9'-H) , 8.29-8.30 (m, 4H, 2x4'-H, 2x6'-H).
¹³ C-NMR (125 MHz, DMSO-d ₆ ): δ = 23.2 (2xC-11 '), 23.7 (C-3, C-4), 35.0 (C-2, C-5), 45.3 (2xC) 1 '), 49.5 (2xC-2'), 59.6 (2xC-6 ''), 61.3 (2xC-10 '), 67.6 (2xC-4''), 70.4 (2xC-2''), 73.1 (2xC -3 ''), 75.2 (2xC-5 ''), 101.1 (2xC-4 '), 102.0 (2xC-1''), 117.6 (2xC-5a'), 122.4 (2xC-9b '), 122.5 ( 2xC-9 '), 123.0 (2xC-7'), 123.2 (2xC-6 '), 126.9 (2xC-8'), 129.3 (2xC-9a '), 141.6 (2xC-3a'), 153.4 (2xC-6 ') 5 '), 170.4 (2xC = O).
MS (ESI): m / z (%) = 1417.4 (4) [3M + 2Na] ²⁺ , 951.3 (100) [M + Na] ⁺ . C ₄₆ H ₅₄ Cl ₂ N ₂ O ₁₄ (929.83). ber .: 951.2844 Found: 951.2839, [M + Na] ⁺ (ESI-HRMS).

Example 13

(-) - 1,7-bis {[(1R, 10S) -1- (10-Chloroethyl) -5-hydroxy-1,2-dihydro-3H-benz [e] indol-3-yl] heptan-1 , 7-dione]} ((-) - (1'R, 10'S) -56c, n = 5)

The phenol (-) - (1R, 10S) -109 (41.3 mg, 119 μmol, 2.0 eq.) Was dissolved in 4 M HCl / ethyl acetate (3.0 mL) and after stirring at room temperature for 1 h the solvent was removed in vacuo. The residue was dried in vacuo for 1 h, dissolved in DMF (3.5 mL), the solution was cooled to 0 ° C. and washed with pimelic acid chloride (66c) (9.9 μL, 11.7 mg, 59.4 μmol, 1.0 eq.) And pyridine (19.2 μL, 18.8 mg, 237 μmol, 4.0 eq.). After stirring for 20 h at room temperature, the reaction mixture was diluted with DMF (3.5 mL) and purified by successive injections (0.4 mL each) by preparative HPLC. The product (-) - (1'S, 10'R) -56c (28.5 mg, 46.0 μmol, 77%) was obtained as a white solid. HPLC (preparative): Pillar: Kromasil 100 C18, 250 × 20 mm, 7 μm Guard column: Kromasil 100 C18, 50 × 20 mm, 5 μm Gradient: Time [min] H ₂ O / MeOH 0 30/70 0-6.0 30/70 → 0/100 6.0-8.5 0/100 8.5-9.5 0/100 → 30/70 9.5-15 30/70 River: 16 mL min ^-1 λ: 254 nm t _R : 9.1 min [Α] 20 / D = -10.7 ° (c = 0.252, DMSO).
UV (CH ₃ CN): λ _max (Ig ε) = 206.5 nm (4,587), 222.5 (4,409) 256.5 (4,950) 264.0 (4,950) 320.0 (4,338).
IR (KBr): ṽ (cm ^-1 ) = 3191, 2930, 1631, 1584, 1522, 1454, 1425, 1394, 1322, 1263, 1157, 1134, 1071, 1024, 860, 757.
¹ H-NMR (600 MHz, DMSO-d ₆ ): δ = 1.50 (m _c , 2H, 4-H ₂ ), 1.62 (d, J = 6.7 Hz, 6H, 2x10'-CH ₃ ), 1.71 (m _c , 4H, 3-H ₂ , 5-H ₂ ), 2.54 (m _C , 4H, 2-H ₂ , 6-H ₂ ), 4.09 (m _C , 2H, 2x1'-H), 4.24-4.25 ( m, 4H, 2x2'-H ₂ ), 4.76 (m _c , 2H, 2x10'-H), 7.30 (t, J = 7.6 Hz, 2H, 2x7'-H), 7.46 (t, J = 7.6 Hz, 2H, 2x8'-H), 7.81 (d, J = 8.5 Hz, 2H, 2x9'-H) , 7.99 (s, 2H, 2x4'-H), 8.09 (d, J = 8.5 Hz, 2H, 2x6'-H), 10.22 (s, 2H, 2xOH).
¹³ C-NMR (125 MHz, DMSO-d ₆ ): δ = 23.2 (2xC-11 '), 24.1 (C-3, C-5), 28.4 (C-4), 35.1 (C-2, C-) 6), 45.2 (2xC-1 '), 49.6 (2xC-2'), 61.5 (2xC-10 '), 99.7 (2xC-4'), 114.6 (2xC-5a '), 121.5 (2xC-9b') , 122.2 (2xC-7 '), 122.5 (2xC-9'), 122.9 (2xC-6 '), 126.6 (2xC-8'), 129.7 (2xC-9a '), 141.8 (2xC-3a'), 153.7 (2xC-5 '), 170.2 (2xC = O).
MS (ESI): m / z (%) = 1261.4 (43) [2M + Na] ⁺ , 641.2 (43) [M + Na] ⁺ , 617.2 (74) [MH] ^- , 581.3 (80) [M-] 2H-Cl] ^- . C ₃₅ H ₃₆ Cl ₂ N ₂ O ₄ (619.58). ber .: 641.1944 Found: 641.1956, [M + Na] ⁺ (ESI-HRMS).

Example 14

(+) - {1,7-bis [(1S, 10R) -1- (-chloroethyl 10) -5-hydroxy-1,2-dihydro-3H-benz [e] indol-3-yl] heptan-1 , 7-dione]} ((+) - (1'S, 10'R) -56c, n = 5)

The phenol (+) - (1S, 10R) -109 (59.4 mg, 171 μmol, 2.0 eq.) Was dissolved in 4 M HCl / ethyl acetate (4.5 mL) and after stirring at room temperature for 1 h, the solvent was removed in vacuo. The residue was dried in vacuo for 1 h, dissolved in DMF (5 mL), the solution was cooled to 0 ° C. and treated with pimelic acid chloride (66c) (14.2 μL, 16.8 mg, 85.4 μmol, 1.0 eq.) And pyridine (27.6 μL, 27.0 mg, 342 μmol, 4.0 eq.). After stirring for 20 h at room temperature, the reaction mixture was diluted with DMF (5 mL) and purified in successive injections (0.4 mL each) by preparative HPLC. The product (+) - (1'S, 10'R) -56c (41.6 mg, 67.1 μmol, 79%) was obtained as a white solid. HPLC (analytical): Pillar: Kromasil 100 C18, 250 × 4.0 mm, 5 μm Gradient: Time [min] H ₂ O / MeOH 0 30/70 0-6 30/70 → 0/100 6-9.5 0/100 9.5-10.0 0/100 → 30/70 10.0-15 30/70 River: 0.8 mL min ^-1 t _R : 9.9 min HPLC (preparative): Pillar: Kromasil 100 C18, 250 × 20 mm, 7 μm Guard column: Kromasil 100 C18, 50 × 20 mm, 5 μm Gradient: Time [min] H ₂ O / MeOH 0 30/70 0-6.0 30/70 → 0/100 6.0-8.5 0/100 8.5-9.5 0/100 → 30/70 9.5-15 30/70 River: 16 mL min ^-1 λ: 254 nm t _R : 9.1 min [Α] 20 / D = + 11.6 ° (c = 0.251, DMSO).
UV (CH ₃ CN): λ _max (Ig ε) = 206.5 nm (4,566), 222.5 (4,401) 256.5 (4,953) 264.0 (4,952) 320.0 (4,337).
IR (KBr): ṽ (cm ^-1 ) = 3184, 2931, 1631, 1584, 1522, 1454, 1424, 1393, 1262, 1157, 1134, 1066, 1024, 860, 756, 632, 413.
¹ H-NMR (600 MHz, CDCl ₃ ): δ = 1.31 (d, J = 6.6 Hz, 6H, 2x10'-CH ₃ ), 1.68-1.77 (m, 4H, 3-H _a , 4-H ₂ , 5-H _a), 2:07 (m _c, 2H, 3-H _b, 5-H _b), 2:44 to 2:50 (m, 4H, H-2x1', 2H _a, 6-H _a), 2.56- 2.61 (m, 2H, 2H _b, 6-H _b), 3.20 (t, J = 9.6 Hz, 2H, 2x2'-H _a), 3.77 (dd, J = 10.4, 1.5 Hz, 2H, 2x2 ' -H _b), 4.18 (m _c, 2H, 2x10'-H), 7.27 (d, J = 8.2 Hz, 2H, 2x9'-H), 7:34 (t, J = 7.4 Hz, 2H, H-2x7' ), 7.44 (t, J = 7.4 Hz, 2H, 2x8'-H), 8.05 (s, 2H, 2x4'-H), 8.29 (d, J = 8.3 Hz, 2H, 2x6'-H), 10.65 ( s, 2H, 2xOH).
¹³ C-NMR (125 MHz, CDCl _3): δ = 23.6 (C-3, C-5), 23.8 (2 × C-11 '), 28.6 (C-4), 36.1 (C-2, C-6) , 45.7 (2xC-1 '), 51.1 (2xC-2'), 60.0 (2xC-10 '), 100.7 (2xC-4'), 114.7 (2xC-5a '), 122.3 (2xC-7'), 122.4 (2 signals) (2xC-9 ', 2xC-9b'), 123.8 (2xC-6 '), 126.0 (2xC-8'), 129.7 (2xC-9a '), 141.3 (2xC-3a'), 154.4 ( 2xC-5 '), 171.7 (2xC = O).
MS (ESI): m / z (%) = 1261.4 (41) [2M + Na] ⁺ , 641.2 (57) [M + Na] ⁺ , 617.2 (52) [MH] ^- , 581.3 (100) [M-] 2H-Cl] ^- . C ₃₅ H 36 Cl ₂ N ₂ O ₄ (619.38). Calc .: 619.2125 Found: 619.2125, [M + H] ⁺ (ESI-HRMS).

Example 15

(-) - {1,7-bis [(1S, 10R) -1- (10-chloroethyl) -5- (β-D-galactopyranosyl) -1,2-dihydro-3H-benz [e] indol-3 -yl] heptane-1,7-dione]} ((-) - (1'S, 10'R) -54c, n = 5)

A mixture (126 mg) of the acetyl protected galactoside (1'S, 10'R) -72c and an ambiguously identified by-product was suspended in absolute methanol (35 mL) and NaOMe in methanol (73 μL of a 5.4 M solution) at 0 ° C , 394 μmol). After warming to room temperature and stirring for 30 min at this temperature, the substance mixture had completely dissolved. Stirring was continued for a further 1.5 h at room temperature, the reaction solution followed by water (3.0 mL) and as long as acidic ion exchanger added (Amberlite IR-120 ^®), until the solution was neutral (pH = 7). The mixture was separated via transfer cannula from the ion exchanger, the same washed with methanol (2 × 6 mL), the organic phases combined and the solvent removed in vacuo. The resulting residue (114 mg) was dissolved in DMF (5 mL) and purified by successive injections (0.4 mL each) by preparative HPLC. The product (-) - (1'S, 10'R) -54c (65.8 mg, 69.7 μmol, 35% over 3 steps) was obtained as a light yellow solid. HPLC (analytical): Pillar: Kromasil 100 C18, 250 × 4.0 mm, 5 μm Gradient: Time [min] H ₂ O / MeOH 0 30/70 0-15 70/30 → 0/100 15-22 0/100 22-23 0/100 → 70/30 23-30 30/70 River: 0.8 mL min ^-1 t _R : 15.3 min HPLC (preparative): Pillar: Kromasil 100 C18, 250 × 20 mm, 7 μm Guard column: Kromasil 100 C18, 50 × 20 mm, 5 μm Gradient: Time [min] H ₂ O / MeOH 0 30/70 0-22.5 70/30 → 17.5 / 82.5 22.5-23.5 17.5 / 82.5 → 70/30 23.5-30 30/70 River: 16 mL min ^-1 λ: 254 nm t _R : 22.8 min [Α] 20 / D = -16.5 ° (c = 0.254, DMSO).
UV (CH ₃ CN): λ _max (Ig ε) = 206.5 nm (4,088), 255.0 (4,275) 262.5 (4,287) 304.5 (3,640) 315.0 (3,700).
IR (KBr): ṽ (cm ^-1 ) = 3406, 2927, 1656, 1580, 1470, 1412, 1074, 758.
¹ H-NMR (600 MHz, DMSO-d ₆ ): δ = 1.50 (m _c , 2H, 4-H ₂ ), 1.65 (d, J = 6.5 Hz, 6H, 2x10'-CH ₃ ), 1.68-1.74 (m, 4H, 3-H ₂ , 5-H ₂ ), 2.57 (m _c , 4H, 2-H ₂ , 6 -H ₂ ), 3.48-3.65 (m, 8H, 2x3 "-H, 2x5 ''-H,2x6''- H _2), 3.75-3.81 (m, 4H, 2x2''- H, 2x4''- H), 4.18 (m _c, 2H, 2x1'-H), 4:27 to 4:33 ( m, 4H, 2x2'-H ₂ ), 4.51 (d, J = 4.5Hz, 2H, 2xOH), 4.55 (m _C , 2H, 2xOH), 4.77-4.86 (m, 4H, 2x10'-H, 2xOH) , 4.93 (d, J = 7.6 Hz, 2H, 2x1 "-H), 5.26 (d, J = 4.9 Hz, 2H, 2xOH), 7.38 (t, J = 7.6 Hz, 2H, 2x7'-H), 7.52 (t, J = 7.5 Hz, 2H, 2x8'-H), 7.89 (d, J = 8.5 Hz, 2H, 2x9'-H), 8.30-8.31 (m, 4H, 2x4'-H, 2x6'- H).
¹³ C-NMR (125 MHz, DMSO-d ₆ ): δ = 23.2 (2xC-11 '), 24.0 (C-3, C-5), 28.3 (C-4), 35.1 (C-2, C-) 6), 45.3 (2xC-1 '), 49.5 (2xC-2'), 59.7 (2xC-6 ''), 61.3 (2xC-10 '), 67.6 (2xC-4''), 70.4 (2xC-2 ''), 73.1 (2xC-3 ''), 75.2 (2xC-5 ''), 101.2 (2xC-4 '), 102.0 (2xC-1''), 117.6 (2xC-5a'), 122.4 (2xC -9b '), 122.5 (2xC-9'), 123.0 (2xC-7 '), 123.2 (2xC-6'), 126.9 (2xC-8 '), 129.4 (2xC-9a'), 141.6 (2xC-3a '), 153.4 (2xC-5'), 170.4 (2xC = O).
MS (ESI): m / z (%) = 965.3 (100) [M + Na] ⁺ , 941.4 (100) [MH] ^- . C ₄₇ H ₅₆ Cl ₂ N ₂ O ₁₄ (943.86). ber .: 965.3001 Found: 965.3036, [M + Na] ⁺ (ESI-HRMS).

Example 16

cytotoxicity

In order to investigate the cytotoxic effect of the newly synthesized substances on adherently growing human tumor cells, a clonogenicity test was carried out, which is based on the HTCFA test and reflects the proliferation capacity of individual cells. As a cell line, the adherently growing human bronchial carcinoma of the line A549 served. The seeding of the tumor cells was carried out from a stock culture, in which they a culture medium (DMEM with the addition of 10% fetal calf serum, 44 mM NaHCO3 and 4 mM glutamine) at 37 ° C and 7.5% CO2 fumigation were kept permanently in air, in concentrations of 10 ² to 10 ⁴ cells in 6 well multi-dishes. After 24 hours, the culture medium was removed, the cells were washed once with the UltraCulture serum-free incubation medium and the cells were subsequently incubated in this medium for 24 hours at ten to twelve different concentrations (three decadic orders) of the substance to be tested. This was previously prepared fresh in a DMSO stock solution and diluted with the incubation medium, so that in the wells ultimately a DMSO concentration of 1% was present. After removing the test substance and another wash with the culture medium, remaining cells were cultured for 12 days in normal culture medium. This was then removed, the clones formed were dried, stained with Löffler's methylene blue and counted macroscopically from a clone size of about 500 microns. The relative cloning rates were determined using the formula given in Figure B.14, where the clone number of the control experiment resulted from untreated cells and was set equal to 100%. The release of the cytotoxic compounds from the prodrugs was carried out by adding 4.0 U / ml of β-D galactosidase. Each series of tests was usually duplicated at least three times to obtain reproducible test results. The results are shown in Table 1. Table 1 compound number IC ₅₀ Selectivity QIC ₅₀ 53a 0.15 pmol / L (m.w.) 146,000 pmol / L (without E.) 970000 53b 5.8 pmol / L (m.w.) 129,000 pmol / L (n / a) 22,000 53c 1.3 pmol / L (m.w.) 180,000 pmol / L (n / a) 140000 54a 150 pmol / L (m.w.) 326 000 pmol / l (no information available) 2200 54b 2860 pmol / L (m.w.) 5,300,000 pmol / L (no information) 1900 54c 260 pmol / L (m.w.) 38,300,000 pmol / L (no data) 150000 55a 0.11 pmol / L 55c 1.0 pmol / L 56a 160 pmol / L 56b 3140 pmol / L 56c 170 pmol / L (m.E.) = with enzyme, (without E.) = without enzyme

Example 17

In Vivo Evaluation of the Compounds of the Invention in an Orthotopic Breast Tumor SCID Mouse Model Using the ADEPT Concept

The female SCID mice are anesthetized with 15 mg / g xylazine prior to planting the tumor cells by peritoneal injection of 75 mg / g ketamine hydrochloride. 1 x 10 ⁶ MDR-MB-231 cells (estrogen-independent human breast cancer cell line) suspended in 5 ml of sterile PBS are then implanted. After implantation, the mice are examined daily for loss of body weight, general condition and tumor formation. On day 21, the animals are randomized into groups of 5 to 12 mice (control, antibody-only, prodrug only and ADEPT therapy). On days 22 and 30, the mice are administered intravenously with PBS (control, prodrug only) or 50 g of a monoclonal anti-human urokinase plasminogen activator receptor (uPAR) antibody conjugated to β-galactosidase, uPAR-β-Gal, in PBS (only antibody enzyme, ADEPT therapy) injected. On days 24, 26, 28, 32, 34 and 36 the mice are injected intravenously with 1% DMSO / NaCl solution (control, antibody-only enzyme) of the compound of the invention 1% ziges DMSO / NaCL solution (only prodrug, ADEPT therapy). The mice are sacrificed on day 38 and the tumors are removed, weighed and fixed in PBS buffer in 4% formalin for 16 hours at room temperature and embedded in paraffin. Tissue sections are assayed for cell proliferation, e.g. B. with an antibody directed against Ki-67. Furthermore, the tumor size is determined by means of computed tomography during the treatment.

Both volume reduction and reduction in metastasis formation can be observed, while no significant impact on weight, leukocytes, hemoglobin and platelets is seen. The tumor growth rate of ADEPT therapy is lower.

QUOTES INCLUDE IN THE DESCRIPTION

This list of the documents listed by the applicant has been generated automatically and is included solely for the better information of the reader. The list is not part of the German patent or utility model application. The DPMA assumes no liability for any errors or omissions.

Cited patent literature

• WO 2007/089149 [0026]
• WO 01/83448 [0026]

Cited non-patent literature

• Mitchell et al., J. Am. Chem. Soc. 1989, 111, 6428-6429 [0015]
• Jia, G., and Lown, W., Bioorg. Med. Chem., 2000, 8, 1607-1617 [0015]

Claims (10)

Compounds of the general structure ALB where A and B are independently selected from the structures I or II

wherein R ^{1 is} selected from a halogen and OSO ₂ R ^u , wherein R ^{u is} selected from optionally substituted C ₁ -C ₆ alkyl, C _1-6 perhaloalkyl, benzyl and phenyl; R ² is selected from hydrogen and optionally substituted C _1-8 alkyl; R ³ , R ^{3 '} , R ⁴ and R ^4' are independently selected from hydrogen and optionally substituted C _1-8 alkyl, wherein two or more of R ² , R ³ , R ^{3 '} , R ⁴ and R ^{4' are} optionally linked together to form one or more optionally substituted carbon cycles or heterocycles; X ² is selected from O, C (R ¹⁴ ) (R ^{14 '} ) and NR ^14' , wherein R ^{14 is} selected from hydrogen and optionally substituted C _1-8 alkyl or C _1-8 acyl and where R ^{14 '} is absent is or is selected from hydrogen and optionally substituted C _1-8 alkyl or C _1-8 acyl; R ⁵ , R ^{5 '} , R ⁶ , R ^6' , R ⁷ and R ^{7 '} are independently selected from H, OH, SH, CF ₃ , CN, C (O) NH ₂ , C (O) H, C (O) OH, halogen, R ^k , SR ^k , S (O) R ^k , S (O) ₂ R ^k , S (O) OR ^k , S (O) ₂ OR ^k , OS (O) R ^k , OS (O) ₂ R ^k , OS (O) OR ^k , OS (O) ₂ R ^k , OR ^k , P (O) (OR ^k ) (OR ^L ), OP (O) (OR ^k ) (OR ^L ), SiR ^k R ^L R ^m , C (O) R ^k , C (O) OR ^k , C (O) N (R ^L ) R ^k , OC (O) R ^k , OC (O) R ^k , OC (O) OR ^k , OC (O) N (R ^k ) R ^L where R ^k , R ^L and R ^{m are} independently selected from H and optionally substituted C _1-4 alkyl, C _1-4 heteroalkyl, C _{3- 7} cycloalkyl, C _3-7 heterocycloalkyl, C _4-12 aryl or C _4-12 heteroaryl groups, two or more of R ^k , R ^L and R ^m optionally together form one or more optionally substituted aliphatic or aromatic carbon cycles or heterocycles; and / or R ^{5 '} and R ^6' and / or R ^{6 '} and R ^7' and / or R ^{7 '} and R ^14' are absent, ie they form a double ring present in the ring between those with the corresponding substituted atoms from, namely R ^{5 '} and R ^6' , and / or R ^{6 '} and R ^7' , and / or R ^{7 '} and R ^14' ; two or more of R ^5, R ^{5 ',} R ^6', R ^{7 ',} R ^14, and R ^14' may optionally be connected together to form one or more optionally substituted aliphatic or aromatic carbon rings or heterocycles; and / or R ⁵ + R ^{5 '} , and / or R ⁶ + R ^6' and / or R ⁷ + R ^{7 '} are independently = O, = S or = NR ¹² , wherein R ^{12 is} selected from H and optionally substituted C _1-6 alkyl; X ₆ is selected from C or N; R _DB is selected from hydrogen and optionally substituted C _1-8 alkyl or optionally substituted C _1-8 acyl; X ₁ is selected from O, S, and NR ¹³ , wherein R ^{13 is} selected from H and optionally substituted C _1-8 alkyl; R is selected from hydrogen or an enzymatically cleavable substrate; a and b are independently selected from 0 and 1; c is selected from 0, 1 and 2; d is selected from 0 or 1; and L is a linking group to the covalent linkage of A and B; and pharmaceutically acceptable salts or pharmaceutically acceptable solvates thereof. The compound of claim 1 wherein L is selected from i) Z- (C (R _CH ) ₂ ) _n -Z ', where n is an integer of 1 to 20, Z and Z' are independently selected from C = O , OC = O, SO ₂ , NR _z , NR _z C = O, C = ONR _z where each R _CH and R _{z are} independently selected from hydrogen or optionally substituted alkyl or optionally substituted acyl, preferably from optionally substituted C ₁ - C ₈ alkyl or optionally substituted C ₁ -C ₈ acyl, or ii) L is an oligopeptide of 1 to 10 amino acids; or pharmaceutically acceptable salts or substrates thereof. A compound according to claim 1 or 2 wherein R is selected from a substrate comprising proteolytic enzymes, plasmin, cathepsin, cathepsin B, beta-glucuronidase, galactosidase, mannosidase, glucosidase, neuramidase, sucrose, maltase, fructosidase, glycosylases, prostate-specific Antigen (PSA), urokinase-type plasminogen activator (u-PA), metalloproteinase, or an enzyme that can be specifically cleaved using targeted enzyme prodrug therapy, such as ADEPT, VDEPT, MDEPT, GDEPT, or PDEPT; or a substituent which can be cleaved or transformed under hypoxic conditions or by reduction by nitroreductase, in particular R is selected from a monosaccharide, disaccharide or oligosaccharide, especially hexoses, pentoses or heptoses optionally as a deoxy derivative or amino derivative and optionally substituted with Halogen, C _1-8 alkyl, C _1-8 acyl, C _1-8 heteroalkyl, C _3-7 cycloalkyl, C _3-7 heterocycloalkyl, C _4-12 aryl or C _4-12 heteroaryl, amino or amide groups, or with amino, amido or carboxy units, which may optionally be substituted by halogen, C _1-8 alkyl, C _1-8 acyl, C _1-8 heteroalkyl, C _3-7 cycloalkyl, C _3-7 heterocycloalkyl, C _{4- 12} aryl or C _4-12 heteroaryl, amino or amide radicals; Dextran, dipeptide, tripeptide, tetrapeptide, oligopeptide, peptidomimetics, or an antibody, or combinations thereof. A compound according to any one of the preceding claims wherein x ^{6 is} nitrogen and / or L is Z- (C (R _CH ) ₂ ) _n -Z 'where n is an integer from 1 to 20, Z and Z' are C = O and each R _CH is independently hydrogen or CH ₃ . A compound according to any one of the preceding claims, wherein a is zero and / or n is an integer of 3, 4 or 5. A compound according to any one of the preceding claims wherein R ⁶ , R ^{6 '} , R ⁷ , R ^7' and R ^{2 are} independently hydrogen or CH ₃ . A compound according to any preceding claim, wherein these are:

wherein R is independently hydrogen or CH ₃ and n is an integer of 3, 4 or 5. A compound according to any one of the preceding claims, wherein A and B are identical. A pharmaceutical composition containing a compound according to any one of claims 1 to 8. Use of a compound according to any one of claims 1 to 8 for the treatment of tumor diseases, especially in mammals.

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