DE102004025731A1 - Antibacterial amide macrocycles III - Google Patents

Abstract

The invention relates to antibacterial amide macrocycles and processes for their preparation, their use for the treatment and / or prophylaxis of diseases and their use for the preparation of medicaments for the treatment and / or prophylaxis of diseases, in particular of bacterial infections.

Description

The This invention relates to antibacterial amide macrocycles and methods for their preparation, their use for treatment and / or prophylaxis of diseases as well as their use for the production of medicines for the treatment and / or prophylaxis of diseases, in particular from bacterial infections.

In WO 03/106480 and WO 04/012816 are antibacterial macrocycles Biphenomycin B type with amide or ester substituents described.

In US 3,452,136 , Dissertation RU Meyer, University of Stuttgart, Germany 1991, Dissertation V. Leitenberger, University of Stuttgart, Germany 1991, Synthesis (1992), (10), 1025-30, J. Chem. Soc., Perkin Trans. 1 (1992), (1), 123-30, J. Chem. Soc., Chem. Commun. (1991), (10), 744, Synthesis (1991), (5), 409-13, J. Chem. Soc., Chem. Commun. (1991), (5), 275-7, J. Antibiot. (1985), 38 (11), 1462-8, J. Antibiot. (1985), 38 (11), 1453-61, the natural product biphenomycin B is described as having antibacterial activity. Partial steps in the synthesis of biphenomycin B are described in Synlett (2003), 4, 522-526.

Chirality (1995), 7 (4), 181-92, J. Antibiot. (1991) 44 (6), 674-7, J. Am. Chem. Soc. (1989), 111 (19), 7323-7, J. Am. Chem. Soc. (1989), 111 (19), 7328-33, J. Org. Chem. (1987), 52 (24), 5435-7, Anal. Biochem. (1987) 165 (1), 108-13, J. Org. Chem. (1985), 50 (8), 1341-2, J. Antibiot. (1993), 46 (3), C-2, J. Antibiot. (1993), 46 (1), 135-40, Synthesis (1992), (12), 1248-54, Appl. Environ. Microbiol. (1992), 58 (12), 3879-82, J. Chem. Soc., Chem. Commun. (1992), (13), 951-3 a structurally related natural product, biphenomycin A, at the Macrocycle has a further substitution with a hydroxy group.

The Natural substances do not correspond to the properties in terms of their properties Requirements for antibacterial drugs. Although structurally different antibacterial on the market effective means available, but it can regularly lead to a development of resistance come. New funds for a good and more effective therapy is therefore desirable.

A Object of the present invention is therefore, new and alternative Compounds with the same or improved antibacterial activity for the treatment of bacterial diseases in humans and animals to disposal to deliver.

Surprisingly, it has been found that certain derivatives of these natural products, wherein the carboxyl group of the natural product is replaced by an amide group containing a basic group, are antibacterially effective against biphenomycin-resistant S. aureus strains (RN4220Bi ^R and T17).

Further in show the derivatives against S. aureus wild-type strains and biphenomycin resistant S. aureus strains an improved spontaneous resistance frequency.

bei denen
R¹ gleich eine Gruppe der Formel

wobei
R⁷ gleich Wasserstoff oder Hydroxy ist,
* die Anknüpfstelle an das Kohlenstoffatom ist,
R² gleich Wasserstoff, Methyl oder Ethyl ist,
R³ gleich eine Gruppe der Formel

ist,
wobei
* die Anknüpfstelle an das Stickstoffatom ist,
R⁸ gleich Wasserstoff, Amino oder Hydroxy ist,
R⁹ gleich Wasserstoff, Methyl oder eine Gruppe der Formel

ist,
worin
* die Anknüpfstelle an das Stickstoffatom ist,
R²⁰ gleich Wasserstoff oder *-(CH₂)_f-NHR²² ist,
worin
R²² gleich Wasserstoff oder Methyl ist
und
f eine Zahl 1, 2 oder 3 ist,
R²¹ gleich Wasserstoff oder Methyl ist,
d eine Zahl 0, 1, 2 oder 3 ist
und
e eine Zahl 1, 2 oder 3 ist,
R¹⁰ gleich Wasserstoff oder Aminoethyl ist,
oder
R⁹ und R¹⁰ bilden zusammen mit dem Stickstoffatom an das sie gebunden sind einen Piperazin-Ring;
R¹¹ und R¹⁵ unabhängig voneinander *-(CH₂)_Z1-OH, *-(CH₂)_Z2-NHR¹⁶, Hydroxycarbonyl, Aminocarbonyl, Hydroxycarbonylmethyl, Aminocarbonylmethyl, *-CONHR¹⁷ oder *-CH₂CONHR¹⁸ sind,
worin
* die Anknüpfstelle an das Kohlenstoffatom ist,
Z1 und Z2 unabhängig voneinander eine Zahl 1, 2 oder 3 sind,
R¹⁶ gleich Wasserstoff oder Methyl ist
und
R¹⁷ und R¹⁸ unabhängig voneinander eine Gruppe der Formel

sind,
worin
* die Anknüpfstelle an das Stickstoffatom ist,
R^8a gleich Wasserstoff, Amino oder Hydroxy ist,
R^9a gleich Wasserstoff, Methyl oder Aminoethyl ist,
R^10a gleich Wasserstoff oder Aminoethyl ist,
oder
R^9a und R^10a bilden zusammen mit dem Stickstoffatom an das sie gebunden sind einen Piperazin-Ring,
R^11a und R^15a unabhängig voneinander *-(CH₂)_Z1a-OH, *-(CH₂)_Z2a-NHR^16a, Hydroxycarbonyl, Aminocarbonyl, Hydroxycarbonylmethyl oder Aminocarbonylmethyl sind,
worin
* die Anknüpfstelle an das Kohlenstoffatom ist,
R^16a gleich Wasserstoff oder Methyl ist
und
Z1a und Z2a unabhängig voneinander eine Zahl 1, 2 oder 3 sind,
R^12a und R^14a unabhängig voneinander Wasserstoff oder Methyl sind,
R^13a gleich Amino oder Hydroxy ist,
ka eine Zahl 0 oder 1 ist,
la, wa, xa und ya unabhängig voneinander eine Zahl 1, 2, 3 oder 4 sind,
R¹² und R¹⁴ unabhängig voneinander Wasserstoff, Methyl oder eine Gruppe der Formel

sind,
worin
* die Anknüpfstelle an das Stickstoffatom ist,
R^20a gleich Wasserstoff oder *-(CH₂)_fa-NHR^22a ist,
worin
R^22a gleich Wasserstoff oder Methyl ist
und
fa eine Zahl 1, 2 oder 3 ist,
R^21a gleich Wasserstoff oder Methyl ist,
da eine Zahl 0, 1, 2 oder 3 ist
und
ea eine Zahl 1, 2 oder 3 ist,
R¹³ gleich Amino oder Hydroxy ist,
R¹⁹ eine Gruppe der Formel

ist,
worin
* die Anknüpfstelle an das Stickstoffatom ist,
R^8b gleich Wasserstoff, Amino oder Hydroxy ist,
R^9b gleich Wasserstoff, Methyl oder Aminoethyl ist,
R^10b gleich Wasserstoff oder Aminoethyl ist,
oder
R^9b und R^10b bilden zusammen mit dem Stickstoffatom an das sie gebunden sind einen Piperazin-Ring,
R^11b und R^15b unabhängig voneinander *-(CH₂)_Z1b-OH, *-(CH₂)_Z2b-NHR^16b, Hydroxycarbonyl, Aminocarbonyl, Hydroxycarbonylmethyl oder Aminocarbonylmethyl sind,
worin
* die Anknüpfstelle an das Kohlenstoffatom ist,
R^16b gleich Wasserstoff oder Methyl ist
und
Z1b und Z2b unabhängig voneinander eine Zahl 1, 2 oder 3 sind,
R^12b und R^14b unabhängig voneinander Wasserstoff oder Methyl sind,
R^13b gleich Amino oder Hydroxy ist,
kb eine Zahl 0 oder 1 ist,
lb, wb, xb und yb unabhängig voneinander eine Zahl 1, 2, 3 oder 4 sind,
k und t unabhängig voneinander eine Zahl 0 oder 1 sind,
l, w, x und y unabhängig voneinander eine Zahl 1, 2, 3 oder 4 sind,
m, r, s und v unabhängig voneinander eine Zahl 1 oder 2 sind,
n, o, p und q unabhängig voneinander eine Zahl 0, 1 oder 2 sind,
u eine Zahl 0, 1, 2 oder 3 ist,

oder y unabhängig voneinander bei l, w, x oder y gleich 3 eine Hydroxy Gruppe tragen kann,
R⁴ gleich Wasserstoff, Halogen, Amino, Hydroxy, Aminocarbonyl, Hydroxycarbonyl, Nitro, Trifluormethyl, Trifluormethoxy, (C₁-C₄)-Alkyl, (C₁-C₄)-Alkoxy oder (C₁-C₆)-Alkylamino ist,
R⁵ gleich Wasserstoff, Halogen, Amino, Hydroxy, Aminocarbonyl, Hydroxycarbonyl, Nitro, Trifluormethyl, Trifluormethoxy, (C₁-C₄)-Alkyl, (C₁-C₄)-Alkoxy oder (C₁-C₆)-Alkylamino ist,
R⁶ gleich Wasserstoff, Methyl oder Ethyl ist,
und ihre Salze, ihre Solvate und die Solvate ihrer Salze.The invention relates to compounds of the formula

at them
R ¹ is a group of the formula

in which
R ⁷ is hydrogen or hydroxy,
* is the point of attachment to the carbon atom,
R ² is hydrogen, methyl or ethyl,
R ³ is a group of the formula

is
in which
* is the point of attachment to the nitrogen atom,
R ⁸ is hydrogen, amino or hydroxy,
R ⁹ is hydrogen, methyl or a group of the formula

is
wherein
* is the point of attachment to the nitrogen atom,
R ²⁰ is hydrogen or * - (CH ₂ ) _f -NHR ²² ,
wherein
R ²² is hydrogen or methyl
and
f is a number 1, 2 or 3,
R ²¹ is hydrogen or methyl,
d is a number 0, 1, 2 or 3
and
e is a number 1, 2 or 3,
R ¹⁰ is hydrogen or aminoethyl,
or
R ⁹ and R ¹⁰ together with the nitrogen atom to which they are attached form a piperazine ring;
R ¹¹ and R ¹⁵ independently of each other * - (CH _{2) Z1} -OH, * - (CH _{2) Z2} -NHR ^16, hydroxycarbonyl, aminocarbonyl, hydroxycarbonylmethyl, aminocarbonylmethyl, * -CONHR ¹⁷ or * -CH ₂ CONHR ^18,
wherein
* is the point of attachment to the carbon atom,
Z1 and Z2 are independently a number 1, 2 or 3,
R ¹⁶ is hydrogen or methyl
and
R ¹⁷ and R ^{18 are} independently a group of the formula

are,
wherein
* is the point of attachment to the nitrogen atom,
R ^8a is hydrogen, amino or hydroxy,
R ^9a is hydrogen, methyl or aminoethyl,
R ^10a is hydrogen or aminoethyl,
or
R ^9a and R ^10a together with the nitrogen atom to which they are attached form a piperazine ring,
R ^11a and R ^{15a are} independently * - (CH ₂ ) _{Z 1a} -OH, * - (CH ₂ ) _Z 2a -NHR ^16a , hydroxycarbonyl, aminocarbonyl, hydroxycarbonylmethyl or aminocarbonylmethyl,
wherein
* is the point of attachment to the carbon atom,
R ^16a is hydrogen or methyl
and
Z1a and Z2a are independently a number 1, 2 or 3,
R ^12a and R ^{14a are} independently hydrogen or methyl,
R ^13a is amino or hydroxy,
ka is a number 0 or 1,
la, wa, xa and ya are independently a number 1, 2, 3 or 4,
R ¹² and R ^{14 are} independently hydrogen, methyl or a group of the formula

are,
wherein
* is the point of attachment to the nitrogen atom,
R ^20a is hydrogen or * - (CH _{2) fa} -NHR ^22a,
wherein
R ^22a is hydrogen or methyl
and
fa is a number 1, 2 or 3,
R ^21a is hydrogen or methyl,
because a number is 0, 1, 2 or 3
and
ea is a number 1, 2 or 3,
R ¹³ is amino or hydroxy,
R ^{19 is} a group of the formula

is
wherein
* is the point of attachment to the nitrogen atom,
R ^8b is hydrogen, amino or hydroxy,
R ^9b is hydrogen, methyl or aminoethyl,
R ^10b is hydrogen or aminoethyl,
or
R ^9b and R ^10b together with the nitrogen atom to which they are attached form a piperazine ring,
R ^11b and R ^{15b are} independently * - (CH ₂ ) _{Z 1b} -OH, * - (CH ₂ ) _Z 2b -NHR ^16b , hydroxycarbonyl, aminocarbonyl, hydroxycarbonylmethyl or aminocarbonylmethyl,
wherein
* is the point of attachment to the carbon atom,
R ^16b is hydrogen or methyl
and
Z1b and Z2b are independently a number 1, 2 or 3,
R ^12b and R ^14b independently of one another are hydrogen or methyl,
R ^13b is amino or hydroxy,
kb is a number 0 or 1,
lb, wb, xb and yb are independently a number 1, 2, 3 or 4,
k and t are independently a number 0 or 1,
l, w, x and y are independently of one another a number 1, 2, 3 or 4,
m, r, s and v are independently of one another a number 1 or 2,
n, o, p and q are independently 0, 1 or 2,
u is a number 0, 1, 2 or 3,

or y independently of l, w, x or y can be a hydroxy group,
R ⁴ is hydrogen, halogen, amino, hydroxy, aminocarbonyl, hydroxycarbonyl, nitro, trifluoromethyl, trifluoromethoxy, (C ₁ -C ₄ ) -alkyl, (C ₁ -C ₄ ) -alkoxy or (C ₁ -C ₆ ) -alkylamino is
R ⁵ is hydrogen, halogen, amino, hydroxy, aminocarbonyl, hydroxycarbonyl, nitro, trifluoromethyl, trifluoromethoxy, (C ₁ -C ₄ ) -alkyl, (C ₁ -C ₄ ) -alkoxy or (C ₁ -C ₆ ) -alkylamino is
R ⁶ is hydrogen, methyl or ethyl,
and their salts, their solvates, and the solvates of their salts.

Compounds according to the invention are the compounds of the formula (I) and their salts, solvates and solvates of the salts, of the compounds of the formula (Ia) mentioned below and of the formula (I) salts, solvates and solvates of the salts as well as those of formula (I) and / or (Ia), hereinafter referred to as embodiment (e) compounds and their salts, solvates and solvates of the salts, as far as in the formula (I ) and / or (Ia), compounds mentioned below are not already salts, solvates and solvates of the salts.

The Compounds of the invention can dependent on of their structure in stereoisomeric forms (enantiomers, diastereomers) exist. The invention therefore includes the enantiomers or Diastereomers and their respective mixtures. From such mixtures enantiomers and / or diastereomers can be the stereoisomer isolate uniform components in a known manner.

Provided the compounds of the invention can occur in tautomeric forms, For example, the present invention encompasses all tautomeric forms.

When Salts are physiologically acceptable in the context of the present invention Salts of the compounds of the invention prefers. Also included are salts that are suitable for pharmaceutical applications themselves are not suitable but for example for insulation or cleaning the compounds of the invention can be used.

physiological Safe salts of the compounds of the invention include acid addition salts of mineral acids, carboxylic acids and sulfonic acids, e.g. Salts of hydrochloric acid, hydrobromic, Sulfuric acid, Phosphoric acid, methane, ethanesulfonic, toluene sulfonic acid, benzenesulfonic, naphthalenedisulfonic, Acetic acid, trifluoroacetic, propionic acid, Lactic acid, Tartaric acid, malic acid, citric acid, fumaric acid, maleic acid and Benzoic acid.

physiological acceptable salts of the compounds of the invention also include Salts more usual Bases, such as by way of example and preferably alkali metal salts (e.g. Sodium and potassium salts), alkaline earth salts (e.g., calcium and magnesium salts) and ammonium salts derived from ammonia or organic amines with 1 to 16 carbon atoms, such as by way of example and preferably ethylamine, Diethylamine, triethylamine, ethyldiisopropylamine, monoethanolamine, Diethanolamine, triethanolamine, dicyclohexylamine, dimethylaminoethanol, procaine, Dibenzylamine, N-methylmorpholine, Arginine, lysine, ethylenediamine and N-methylpiperidine.

When In the context of the invention, solvates are those forms of the compounds according to the invention, which in solid or liquid Condition by coordination with solvent molecules a complex form. Hydrates are a special form of solvates in which the coordination with water takes place.

Unless otherwise specified, in the context of the present invention, the substituents have the following meaning:
Alkyl per se and "Alk" and "alkyl" in alkoxy and alkylamino are a linear or branched alkyl radical having usually 1 to 6, preferably 1 to 4, particularly preferably 1 to 3 carbon atoms, by way of example and preferably methyl, ethyl, n-propyl, isopropyl, tert -butyl, n -pentyl and n -hexyl.
Alkoxy is by way of example and preferably methoxy, ethoxy, n-propoxy, isopropoxy, tert-butoxy, n-pentoxy and n-hexoxy.
Alkylamino represents an alkylamino radical having one or two (independently selected) alkyl substituents. (C ₁ -C ₃ ) -Alkylamino is, for example, a monoalkylamino radical having 1 to 3 carbon atoms or a dialkylamino radical having in each case 1 to 3 carbon atoms per alkyl substituent. The following may be mentioned by way of example and by way of preference: methylamino, ethylamino, n-propylamino, isopropylamino, tert-butylamino, n-pentylamino, n-hexylamino, N, N-dimethylamino, N, N-diethylamino, N-ethyl-N-methylamino, N- Methyl-Nn-propylamino, N-isopropyl-Nn-propylamino, N-tert-butyl-N-methylamino, N-ethyl-Nn-pentylamino and Nn-hexyl-N-methylamino.
Halogen is fluorine, chlorine, bromine and iodine.

A symbol * in the formula of a group indicates the point of attachment of the group. The end point of the line, next to each of which a *, does not stand for a carbon atom or a CH ₂ group but is part of the bond to the atom to which the group is attached.

In the formulas of the groups which R ^{1 may} stand for, the endpoint of the line next to each of which * is not a carbon atom or a CH ₂ group but is part of the bond to the carbon atom to which R ^{1 is} attached is.

In the formulas of the groups for which R ^{3 can} stand, the endpoint of the line stands next to each a * is not a carbon atom or a CH ₂ group but is part of the bond to the nitrogen atom to which R ^{3 is} attached. R ³ is therefore for example 2-aminoethyl in the case of k = 0, l = 1, R ⁹ = H and R ¹⁰ = H, 3-amino-2-hydroxypropyl in the case of k = 1, R ⁸ = OH, l = 1, R ⁹ = H and R ¹⁰ = H, piperidin-4-yl-methyl in the case of q = 1 and r = 1 or piperidin-4-yl in the case of q = 0 and r = 1.

entsprechen, worin R¹ bis R⁶ die gleiche Bedeutung wie in Formel (I) haben,
und ihre Salze, ihre Solvate und die Solvate ihrer Salze.Preferred in the context of the present invention are compounds which are of the formula

in which R ¹ to R ^{6 have} the same meaning as in formula (I),
and their salts, their solvates, and the solvates of their salts.

wobei
R⁷ gleich Wasserstoff oder Hydroxy ist,
* die Anknüpfstelle an das Kohlenstoffatom ist,
R² gleich Wasserstoff oder Methyl ist,
R³ gleich eine Gruppe der Formel

ist,
wobei
* die Anknüpfstelle an das Stickstoffatom ist,
R⁸ gleich Wasserstoff, Amino oder Hydroxy ist,
R⁹ gleich Wasserstoff, Methyl oder eine Gruppe der Formel

ist,
worin
* die Anknüpfstelle an das Stickstoffatom ist,
R²⁰ gleich Wasserstoff oder *-(CH₂)_f-NHR²² ist,
worin
R²² gleich Wasserstoff ist
und
f eine Zahl 1, 2 oder 3 ist,
R²¹ gleich Wasserstoff ist,
d eine Zahl 0 ist
und
e eine Zahl 1, 2 oder 3 ist,
R¹⁰ gleich Wasserstoff oder Aminoethyl ist,
R¹¹ gleich *-(CH₂)_Z1-OH, *-(CH₂)_Z2-NHR¹⁶, Hydroxycarbonyl, Aminocarbonyl, Hydroxycarbonylmethyl, Aminocarbonyl-methyl, *-CONHR¹⁷ oder *-CH₂CONHR¹⁸ ist,
worin
* die Anknüpfstelle an das Kohlenstoffatom ist,
Z1 und Z2 unabhängig voneinander eine Zahl 1, 2 oder 3 sind,
R¹⁶ gleich Wasserstoff oder Methyl ist
und
R¹⁷ und R¹⁸ unabhängig voneinander eine Gruppe der Formel

sind,
worin
* die Anknüpfstelle an das Stickstoffatom ist,
R^8a gleich Wasserstoff, Amino oder Hydroxy ist,
R^9a gleich Wasserstoff, Methyl oder Aminoethyl ist,
R^10a gleich Wasserstoff oder Aminoethyl ist,
oder
R^9a und R^{10 a} bilden zusammen mit dem Stickstoffatom an das sie gebunden sind einen Piperazin-Ring,
R^{11 a} und R^15a unabhängig voneinander *-(CH₂)_Z1a-OH, *-(CH₂)_Z2a-NHR^16a, Hydroxycarbonyl, Aminocarbonyl, Hydroxycarbonylmethyl oder Aminocarbonylmethyl sind,
worin
* die Anknüpfstelle an das Kohlenstoffatom ist,
R^{1 6a} gleich Wasserstoff oder Methyl ist
und
Z1a und Z2a unabhängig voneinander eine Zahl 1, 2 oder 3 sind,
R^{12 a} und R^14a unabhängig voneinander Wasserstoff oder Methyl sind,
R^13a gleich Amino oder Hydroxy ist,
ka eine Zahl 0 oder 1 ist,
la, wa, xa und ya unabhängig voneinander eine Zahl 1, 2, 3 oder 4 sind,
R¹² gleich Wasserstoff oder Methyl ist,
k gleich eine Zahl 0 oder 1 ist,
l und w unabhängig voneinander eine Zahl 1, 2, 3 oder 4 sind,

oder w unabhängig voneinander bei 1 oder w gleich 3 eine Hydroxy-Gruppe tragen kann,
R⁴ gleich Hydroxy ist,
R⁵ gleich Wasserstoff ist,
R⁶ gleich Wasserstoff ist,
und ihre Salze, ihre Solvate und die Solvate ihrer Salze.Preferred within the scope of the present invention are compounds according to the invention in which
R ¹ is a group of the formula

in which
R ⁷ is hydrogen or hydroxy,
* is the point of attachment to the carbon atom,
R ² is hydrogen or methyl,
R ³ is a group of the formula

is
in which
* is the point of attachment to the nitrogen atom,
R ⁸ is hydrogen, amino or hydroxy,
R ⁹ is hydrogen, methyl or a group of the formula

is
wherein
* is the point of attachment to the nitrogen atom,
R ²⁰ is hydrogen or * - (CH ₂ ) _f -NHR ²² ,
wherein
R ²² is hydrogen
and
f is a number 1, 2 or 3,
R ²¹ is hydrogen,
d is a number 0
and
e is a number 1, 2 or 3,
R ¹⁰ is hydrogen or aminoethyl,
R ¹¹ is * - (CH _{2) Z1} -OH, * - (CH _{2) Z2} -NHR ^16, hydroxycarbonyl, aminocarbonyl, hydroxycarbonylmethyl, aminocarbonyl-methyl, * -CONHR ¹⁷ or * -CH ₂ CONHR ^18,
wherein
* is the point of attachment to the carbon atom,
Z1 and Z2 are independently a number 1, 2 or 3,
R ¹⁶ is hydrogen or methyl
and
R ¹⁷ and R ^{18 are} independently a group of the formula

are,
wherein
* is the point of attachment to the nitrogen atom,
R ^8a is hydrogen, amino or hydroxy,
R ^9a is hydrogen, methyl or aminoethyl,
R ^10a is hydrogen or aminoethyl,
or
R ^9a and R ^{10 a} together with the nitrogen atom to which they are attached form a piperazine ring,
R ^{11 a} and R ^{15a are} independently * - (CH ₂ ) _{Z 1a} -OH, * - (CH ₂ ) _Z 2a -NHR ^16a , hydroxycarbonyl, aminocarbonyl, hydroxycarbonylmethyl or aminocarbonylmethyl,
wherein
* is the point of attachment to the carbon atom,
R ^{1 6a} is hydrogen or methyl
and
Z1a and Z2a are independently a number 1, 2 or 3,
R ^{12 a} and R ^{14a are} independently hydrogen or methyl,
R ^13a is amino or hydroxy,
ka is a number 0 or 1,
la, wa, xa and ya are independently a number 1, 2, 3 or 4,
R ¹² is hydrogen or methyl,
k is a number 0 or 1,
l and w are independently of one another a number 1, 2, 3 or 4,

or w independently of one another can carry a hydroxyl group at 1 or w equals 3,
R ⁴ is hydroxy,
R ⁵ is hydrogen,
R ⁶ is hydrogen,
and their salts, their solvates, and the solvates of their salts.

• [A] Verbindungen der Formel
  
  worin R¹, R², R⁴, R⁵ und R⁶ die oben angegebene Bedeutung haben und boc gleich tert-Butoxycarbonyl ist, in einem zweistufigen Verfahren zunächst in Gegenwart von einem oder mehreren Dehydratisierungsreagenzien mit Verbindungen der Formel H₂NR³ (III),worin R³ die oben angegebene Bedeutung hat, und anschließend mit einer Säure umgesetzt werden, oder
• [B] Verbindungen der Formel
  
  worin R¹, R², R⁴, R⁵ und R⁶ die oben angegebene Bedeutung haben und Z gleich Benzyloxycarbonyl ist, in einem zweistufigen Verfahren zunächst in Gegenwart von einem oder mehreren Dehydratisierungsreagenzien mit Verbindungen der Formel H₂NR³ (III),worin R³ die oben angegebene Bedeutung hat, und anschließend mit einer Säure oder durch Hydrogenolyse umgesetzt werden, oder
• [C] Verbindungen der Formel
  
  worin R¹, R², R⁴, R⁵ und R⁶ die oben angegebene Bedeutung haben und Z gleich Benzyloxycarbonyl ist, und R²³ gleich Benzyl, Methyl oder Ethyl ist, in einem zweistufigen Verfahren zunächst in Gegenwart von Kaliumcyanid mit Verbindungen der Formel H₂NR³ (III),worin R³ die oben angegebene Bedeutung hat, und anschließend mit einer Säure oder durch Hydrogenolyse umgesetzt werden.

The invention furthermore relates to a process for the preparation of the compounds of the formula (I) or their salts, their solvates or the solvates of their salts, according to processes

• [A] Compounds of the formula
  
  wherein R ¹ , R ² , R ⁴ , R ⁵ and R ^{6 have} the abovementioned meaning and boc is tert-butoxycarbonyl, in a two-stage process, first in the presence of one or more dehydrating reagents with compounds of the formula H ₂ NR ³ (III), wherein R ^{3 has} the meaning given above, and then reacted with an acid, or
• [B] Compounds of the formula
  
  wherein R ¹ , R ² , R ⁴ , R ⁵ and R ^{6 have} the abovementioned meaning and Z is benzyloxycarbonyl, in a two-stage process, first in the presence of one or more dehydrating reagents with compounds of the formula H ₂ NR ³ (III), wherein R ^{3 has} the meaning given above, and then reacted with an acid or by hydrogenolysis, or
• [C] Compounds of the formula
  
  wherein R ¹ , R ² , R ⁴ , R ⁵ and R ⁶ are as defined above and Z is benzyloxycarbonyl, and R ²³ is benzyl, methyl or ethyl, in a two-step process, first in the presence of potassium cyanide with compounds of the formula H ₂ NR ³ (III), wherein R ^{3 has} the meaning given above, and then reacted with an acid or by hydrogenolysis.

The Free base of the salts can be obtained, for example, by chromatography a reversed phase column with an acetonitrile-water gradient with the addition of a base especially by using a RP18 Phenomenex Luna C18 (2) column and diethylamine as the base.

Another The invention relates to a process for the preparation of the compounds of formula (I) or its solvates according to claim 1, in which salts the compounds or solvates of the salts of the compounds by chromatography are added with the addition of a base in the compounds or their solvates.

The hydroxy group on R ⁷ is optionally protected during the reaction of the compounds of formulas (IX) and (XI) with a tert-butyldimethylsilyl group and is cleaved off in a subsequent reaction steps under Desilylierungsbedingungen with.

Reactive functionalities in the radicals R ¹ and R ³ of compounds of the formulas (II), (III), (IV) and (VI) are already protected with introduced into the synthesis, preference is given to acid-labile protective groups (eg boc or z). After conversion into compounds of the formula (I), the protective groups can be eliminated by deprotection reaction. This is done by standard methods of protecting group chemistry. Deprotection reactions under acidic conditions or by hydrogenolysis are preferred.

Reactive functionalities in the radical R ¹ of compounds of the formulas (VII), (VIII), (IX) and (XI) are already protected in the synthesis, preference is given to acid-labile protective groups (eg boc or z). These protecting groups are cleaved in one of the subsequent reaction steps by deprotection reaction. This is done by standard methods of protecting group chemistry. Deprotection reactions under acidic conditions or by hydrogenolysis are preferred.

The Implementation of the first stage of the processes [A] and [B] takes place in Generally in inert solvents, optionally in the presence of a base, preferably in a temperature range from 0 ° C up to 40 ° C at normal pressure.

When Dehydrating reagents are suitable here, for example, carbodiimides such as. N, N'-diethyl, N, N'-dipropyl, N, N'-diisopropyl, N, N'-dicyclohexylcarbodiimide, N- (3-dimethylaminoisopropyl) -N'-ethylcarbodiimide hydrochloride (EDC), N-cyclohexylcarbodiimide-N'-propyloxymethyl-polystyrene (PS carbodiimide) or carbonyl compounds such as carbonyldiimidazole, or 1,2-oxazolium compounds such as 2-ethyl-5-phenyl-1,2-oxazolium-3-sulfate or 2-tert-butyl-5-methylisoxazolium perchlorate, or acylamino compounds such as 2-ethoxy-1-ethoxycarbonyl-1,2-dihydroquinoline, or propanephosphonic anhydride, or isobutyl chloroformate, or bis (2-oxo-3-oxazolidinyl) -phosphoryl chloride or Benzotriazolyloxy-tri (dimethylamino) phosphonium hexafluorophosphate, or O- (benzotriazol-1-yl) -N, N, N ', N'-tetra-methyluronium hexafluorophosphate (HBTU), 2- (2-oxo-1- (2H) -pyridyl) -1,1,3,3-tetramethyluronium tetrafluoroborate (TPTU) or O- (7-azabenzotriazol-1-yl) -N, N, N ', N'-tetramethyluronium hexafluorophosphate (HATU), or 1-hydroxybenzotriazole (HOBt), or benzotriazol-1-yloxytris (dimethylamino) phosphonium hexafluoro-phosphate (BOP), or mixtures of these, or mixtures of these together with bases.

bases For example, alkali metal carbonates, e.g. Sodium or potassium carbonate, or bicarbonate, or organic bases such as trialkylamines e.g. Triethylamine, N-methylmorpholine, N-methylpiperidine, 4-dimethylaminopyridine or diisopropylethylamine.

Preferably the condensation with HATU in the presence of a base, in particular Diisopropylethylamine, performed.

inert solvent For example, halogenated hydrocarbons such as dichloromethane or trichloromethane, hydrocarbon, such as benzene, or nitromethane, Dioxane, dimethylformamide or acetonitrile. It is also possible to mix the solvent use. Particularly preferred is dimethylformamide.

The implementation of the first step of the process [C] is generally carried out with the compound of For mel (III) as solvent, preferably in a temperature range from 0 ° C to 40 ° C at atmospheric pressure.

The Reaction with an acid in the second step of the processes [A], [B] and [C] is preferably carried out in a temperature range of 0 ° C up to 40 ° C at normal pressure.

When acids Hydrogen chloride in dioxane, hydrogen bromide are suitable here in acetic acid or trifluoroacetic acid in methylene chloride.

The Hydrogenolysis in the second stage of the processes [B] and [C] is carried out generally in a solvent in the presence of hydrogen and palladium on carbon, preferred in a temperature range of 0 ° C up to 40 ° C at normal pressure.

solvent are, for example, alcohols such as methanol, ethanol, n-propanol or Iso-propanol, in a mixture with water and glacial acetic acid, is preferred is a mixture of ethanol, water and glacial acetic acid.

The Compounds of the formula (III) are known or can be analogous known methods are produced.

worin R¹, R², R⁴, R⁵ und R⁶ die oben angegebene Bedeutung haben,
mit Di-(tert-butyl)-dicarbonat in Gegenwart einer Base umgesetzt werden.The compounds of the formula (II) are known or can be prepared by reacting compounds of the formula

in which R ¹ , R ² , R ⁴ , R ⁵ and R ^{6 have} the abovementioned meaning,
be reacted with di (tert-butyl) dicarbonate in the presence of a base.

The Reaction is generally carried out in a solvent, preferably in a temperature range of 0 ° C up to 40 ° C at normal pressure.

bases For example, alkali hydroxides such as sodium or potassium hydroxide, or alkali carbonates such as cesium carbonate, Sodium or potassium carbonate, or other bases such as DBU, triethylamine or diisopropylethylamine, preferred is sodium hydroxide or sodium carbonate.

solvent For example, halogenated hydrocarbons such as methylene chloride or 1,2-dichloroethane, alcohols, such as methanol, ethanol or isopropanol, or water.

Preferably React the reaction with sodium hydroxide in water or sodium carbonate carried out in methanol.

worin R¹, R², R⁴, R⁵ und R⁶ die oben angegebene Bedeutung haben, und
R²³ gleich Benzyl, Methyl oder Ethyl ist,
mit einer Säure oder durch Hydrogenolyse, wie für die zweite Stufe des Verfahrens [B] beschrieben, gegebenenfalls durch anschließende Umsetzung mit einer Base zur Verseifung des Methyl- oder Ethylesters, umgesetzt werden.The compounds of the formula (V) are known or can be prepared by reacting compounds of the formula

wherein R ¹ , R ² , R ⁴ , R ⁵ and R ^{6 have} the meaning given above, and
R ²³ is benzyl, methyl or ethyl,
with an acid or by hydrogenolysis as described for the second step of process [B], optionally followed by reaction with a base to saponify the methyl or ethyl ester.

The Saponification can be done, for example, as in the implementation of Compounds of formula (VI) to compounds of formula (IV) described.

The Compounds of formula (IV) are known or can be prepared by: in compounds of formula (VI) the benzyl, methyl or ethyl esters is saponified.

The Reaction generally takes place in a solvent, in the presence a base, preferably in a temperature range from 0 ° C to 40 ° C at atmospheric pressure.

bases For example, alkali metal hydroxides such as lithium, sodium or Potassium hydroxide, preferably lithium hydroxide.

solvent For example, halogenated hydrocarbons such as dichloromethane or trichloromethane, ethers such as tetrahydrofuran or dioxane, or Alcohols such as methanol, ethanol or isopropanol, or dimethylformamide. It is also possible Mixtures of solvents or mixtures of the solvents to use with water. Particularly preferred are tetrahydrofuran or a mixture of methanol and water.

worin R¹, R², R⁴, R⁵, R⁶ und R²³ die oben angegebene Bedeutung haben,
in der ersten Stufe mit Säuren, wie für die zweite Stufe der Verfahren [A] und [B] beschrieben, und in der zweiten Stufe mit Basen umgesetzt werden.The compounds of the formula (VI) are known or can be prepared by reacting compounds of the formula

in which R ¹ , R ² , R ⁴ , R ⁵ , R ⁶ and R ^{23 have} the abovementioned meaning,
in the first stage with acids as described for the second stage of processes [A] and [B], and in the second stage with bases.

In In the second step, the reaction with bases generally takes place in a solvent, preferably in a temperature range from 0 ° C to 40 ° C at atmospheric pressure.

bases For example, alkali hydroxides such as sodium or potassium hydroxide, or alkali carbonates such as cesium carbonate, Sodium or potassium carbonate, or other bases such as DBU, triethylamine or diisopropylethylamine, preferred is triethylamine.

solvent For example, halohydrocarbons such as chloroform, methylene chloride or 1,2-dichloroethane, or tetrahydrofuran, or mixtures of the solvents, preferred is methylene chloride or tetrahydrofuran.

worin R¹, R², R⁴, R⁵, R⁶ und R²³ die oben angegebene Bedeutung haben,
mit Pentafluorphenol in Gegenwart von Dehydratisierungsreagenzien, wie für die erste Stufe der Verfahren [A] und [B] beschrieben, umgesetzt werden.The compounds of formula (VII) are known or can be prepared by reacting compounds of the formula

in which R ¹ , R ² , R ⁴ , R ⁵ , R ⁶ and R ^{23 have} the abovementioned meaning,
with pentafluorophenol in the presence of dehydrating reagents as described for the first step of processes [A] and [B].

The Reaction is preferably carried out with DMAP and EDC in dichloromethane in a temperature range of -40 ° C up to 40 ° C at normal pressure.

worin R¹, R², R⁴, R⁵, R⁶ und R²³ die oben angegebene Bedeutung haben,
mit Fluorid, insbesondere mit Tetrabutylammoniumfluorid, umgesetzt werden.The compounds of the formula (VIII) are known or can be prepared by reacting compounds of the formula

in which R ¹ , R ² , R ⁴ , R ⁵ , R ⁶ and R ^{23 have} the abovementioned meaning,
be reacted with fluoride, in particular with tetrabutylammonium fluoride.

The Reaction is generally carried out in a solvent, preferably in a temperature range of -10 ° C up to 30 ° C at normal pressure.

inert solvent are, for example, halogenated hydrocarbons such as dichloromethane, or hydrocarbons such as benzene or toluene, or ethers such as tetrahydrofuran or dioxane, or dimethylformamide. It is also possible to mix the solvent use. Preferred solvents are tetrahydrofuran and dimethylformamide.

worin R², R⁴, R⁵ und R²³ die oben angegebene Bedeutung haben,
mit Verbindungen der Formel

worin R¹ und R⁶ die oben angegebene Bedeutung haben,
in Gegenwart von Dehydratisierungsreagenzien, wie für die erste Stufe der Verfahren [A] und [B] beschrieben, umgesetzt werden.The compounds of formula (IX) are known or can be prepared by reacting compounds of the formula

wherein R ² , R ⁴ , R ⁵ and R ^{23 have} the abovementioned meaning,
with compounds of the formula

in which R ¹ and R ^{6 have} the abovementioned meaning,
in the presence of dehydrating reagents as described for the first step of processes [A] and [B].

The Compounds of the formula (X) are known or can be prepared analogously to those in the examples be prepared described methods.

The Compounds of the formula (XI) are known or can be prepared analogously to known processes getting produced.

The preparation of the compounds according to the invention can be illustrated by the following synthesis scheme: Synthesis of biphenomycin A derivatives using the example of (7R, 8S, 11S, 14S) -14-amino-N- (2-aminoethyl) -11 - [(2R) - 3-amino-2-hydroxypropyl] -5,7,17-trihydroxy-10,13-dioxo-9,12-diazatricyclo [14.3.1.1 ^2,6 ] henicosa-1 (20), 2 (21), 3, 5,16,18-hexaene-8-carboxamide trihydrochloride

The Compounds of the invention show an unpredictable, valuable pharmacological and pharmacokinetic spectrum of activity.

she are therefore suitable for use as a medicament for treatment and / or prophylaxis of diseases in humans and animals.

The Compounds of the invention can due to their pharmacological properties alone or in combination with other active ingredients for the treatment and / or prophylaxis of infectious diseases, especially of bacterial infections.

For example, local and / or systemic diseases caused and / or prevented by the following pathogens or by mixtures of the following pathogens can be treated and / or prevented:
Gram-positive cocci, eg staphylococci (Staph aureus, Staph epidermidis) and streptococci (Strept agalactiae, Strept faecalis, Strept pneumoniae, Strept pyogenes); Gram-negative cocci (Neisseria gonorrhoeae) as well as Gram-negative rods such as Enterobacteriaceae, eg Escherichia coli, Hemophilus influenzae, Citrobacter (Citrobanthusii, Citrob. divernis), Salmonella and Shigella; also Klebsiella (Klebs. pneumoniae, Klebs. oxytocy), Enterobacter (Ent. aerogenes, Ent. agglomerans), Hafnia, Serratia (Serr. marcescens), Proteus (Pr. mirabilis, Pr. rettgeri, Pr. vulgaris), Providencia, Yersinia , as well as the genus Acinetobacter. In addition, the antibacterial spectrum includes the genus Pseudomonas (Ps. Aeruginosa, Ps. Maltophilia) and strictly anaerobic bacteria such as Bacteroides fragilis, members of the genus Peptococcus, Peptostreptococcus and the genus Clostridium; also mycoplasma (M. pneumoniae, M. hominis, M. urealyticum) as well as mycobacteria, eg Mycobacterium tuberculosis.

The above enumeration of pathogens is merely exemplary and by no means limiting. Examples of diseases which are caused by the named pathogens or mixed infections and which can be prevented, ameliorated or cured by the topically applicable preparations according to the invention are:
Infectious diseases in humans such. As septic infections, bone and joint infections, skin infections, postoperative wound infections, abscesses, phlegmon, wound infections, infected burns, burns, infections in the mouth, infections after dental surgery, septic arthritis, mastitis, tonsillitis, genital infections and eye infections.

Apart from humans, bacterial infections can also be treated in other species. Examples include:
Pig: coli-diarrhea, enterotoxemia, sepsis, dysentery, salmonellosis, metritis-mastitis-agalactiae syndrome, mastitis;
Ruminants (cattle, sheep, goats): diarrhea, sepsis, bronchopneumonia, salmonellosis, pasteurellosis, mycoplasmosis, genital infections;
Horse: bronchopneumonia, foal disease, puerperal and postpuerperal infections, salmonellosis;
Dog and cat: bronchopneumonia, diarrhea, dermatitis, otitis, urinary tract infections, prostatitis;
Poultry (chicken, turkey, quail, pigeon, ornamental birds and others): mycoplasmosis, E. coli infections, chronic respiratory diseases, salmonellosis, pasteurellosis, psittacosis.

As well can bacterial diseases in the rearing and keeping of and ornamental fish are treated, being the antibacterial Spectrum over the aforementioned pathogens point to other pathogens such as e.g. Pasteurella, Brucella, Campylobacter, Listeria, Erysipelothris, Corynebacteria, Borellia, Treponema, Nocardia, Rikettsie, Yersinia, extended.

Another The present invention is the use of the compounds of the invention for the treatment and / or prophylaxis of diseases, preferably of bacterial diseases, especially of bacterial infections.

Another The present invention is the use of the compounds of the invention for the treatment and / or prophylaxis of diseases, in particular the aforementioned diseases.

Another The present invention is the use of the compounds of the invention for the manufacture of a medicament for the treatment and / or prophylaxis of diseases, in particular the aforementioned diseases.

Another The subject of the present invention is a method of treatment and / or prophylaxis of diseases, in particular those mentioned above Diseases using an antibacterially effective amount the compounds of the invention.

The Compounds of the invention can act systemically and / or locally. For this purpose they can be applied in a suitable manner, e.g. oral, parenteral, pulmonary, nasal, sublingual, lingual, buccal, rectal, dermal, transdermal, conjunctival, otic or as an implant or stent.

For this Application routes can the compounds of the invention be administered in suitable administration forms.

For the oral Application are functioning according to the prior art rapidly and / or modifies the compounds according to the invention donating Application forms containing the compounds of the invention in crystalline and / or amorphized and / or dissolved form, e.g. Tablets (uncoated or coated Tablets, for example, with enteric or delayed dissolving or insoluble coatings that control the release of the compound of the invention), in the oral cavity rapidly disintegrating tablets or films / wafers, films / lyophilisates, Capsules (for example hard or soft gelatine capsules), dragees, Granules, pellets, powders, emulsions, suspensions, aerosols or Solutions.

The Parenteral administration can bypass a resorption step happen (e.g., intravenously, intraarterial, intracardiac, intraspinal or intralumbar) or involving absorption (e.g., intramuscular, subcutaneous, intracutaneous, percutaneous or intraperitoneal). For parenteral administration are suitable as application forms u.a. Injection and infusion preparations in the form of solutions, Suspensions, emulsions, lyophilisates or sterile powders.

For the others Application routes are suitable, e.g. Inhalation medicines (i.a. Powder inhalers, nebulizers), nasal drops, solutions, sprays; lingual, sublingual or buccally applied tablets, films / wafers or capsules, Suppositories, ear or eye preparations, vaginal capsules, aqueous Suspensions (lotions, shake mixtures), lipophilic suspensions, ointments, creams, transdermal therapeutic Systems (such as patches), milk, pastes, foams, Implants or stents.

The Compounds of the invention can in the cited Application forms are transferred. This can be done in a conventional manner by mixing with inert, Non-toxic, pharmaceutically suitable excipients happen. Among these adjuvants include et al excipients (for example, microcrystalline cellulose, lactose, mannitol), solvent (e.g., liquid Polyethylene glycols), emulsifiers and dispersing or wetting agents (For example, sodium dodecyl sulfate, Polyoxysorbitanoleat), binder (For example, polyvinylpyrrolidone), synthetic and natural polymers (for example, albumin), stabilizers (e.g., antioxidants such as for example ascorbic acid), Dyes (e.g., inorganic pigments such as iron oxides) and flavor and / or scent remedies.

Another The present invention relates to medicaments which are at least a compound of the invention, usually together with one or more inert, non-toxic, pharmaceutical contain suitable excipients, as well as their use to the previously mentioned purposes.

in the In general, it has proven to be beneficial in parenteral Application amounts of about 5 to 250 mg / kg body weight per 24 h to achieve to deliver effective results. For oral administration, the amount is about 5 to 100 mg / kg of body weight every 24 h.

Nevertheless it may be necessary, if necessary, of the quantities mentioned to deviate, depending on of body weight, Route of administration, individual behavior towards the active substance, type of Preparation and time or interval to which the application he follows. So it can in some cases be sufficient, with less than the aforementioned minimum quantity to get along while in other cases exceeded the mentioned upper limit must become. In case of application of larger quantities it may be recommended be to distribute these in several single doses throughout the day.

The Percentages in the following tests and examples are provided not stated otherwise, weight percentages; Parts are parts by weight. Solvent ratios, dilution ratios and concentration data of liquid / liquid solutions each on the volume.

A. Examples

Used abbreviations:

Aloc

allyloxycarbonyl

aq.

aqueous

Equiv.

equivalent to

Bn

benzyl

Boc

tert-butoxycarbonyl

CDCl ₃

chloroform

CH

cyclohexane

d

Doublet (in ¹ H-NMR)

dd

Doublet of doublet

DCM

dichloromethane

DCC

dicyclohexylcarbodiimide

DIC

diisopropylcarbodiimide

DIPEA

diisopropylethylamine

DMSO

dimethyl sulfoxide

DMAP

4-N, N-dimethylaminopyridine

DMF

dimethylformamide

d. Th.

the theory

EDC

N '- (3-dimethylaminopropyl) -N-ethylcarbodiimide × HCl

EE

Ethyl acetate (ethyl acetate)

IT I

Electrospray ionization (in MS)

ges.

saturated

HATU

O- (7-azabenzotriazol-1-yl) -N, N, N ', N'-tetramethyluronium hexafluorophosphate

HBTU

O- (benzotriazol-1-yl) -N, N, N ', N'-tetramethyluronium hexafluorophosphate

HOBt

1-hydroxy-1H-benzotriazole × H ₂ O

H

Hour (s)

HPLC

High pressure, high performance liquid chromatography

conc.

concentrated

LC-MS

Liquid chromatography-coupled mass spectroscopy

m

Multiplet (in ¹ H-NMR)

min

minutes

MS

mass spectroscopy

MeOH

methanol

NMR

Nuclear Magnetic Resonance Spectroscopy

MTBE

Methyl tert-butyl ether

Pd / C

Palladium / carbon

proc.

percent

q

Quartet (in ¹ H-NMR)

R _f

Retention index (at DC)

RT

room temperature

R _t

Retention time (at HPLC)

s

Singlet (in ¹ H-NMR)

t

Triplet (in ¹ H-NMR)

TBS

tert-butyldimethylsilyl

THF

tetrahydrofuran

TMSE

2- (trimethylsilyl) ethyl

TPTU

2- (2-oxo-1 (2H) -pyridyl) -1,1,3,3-tetramethyluronium tetrafluoroborate

Z

benzyloxycarbonyl

LC-MS and HPLC methods:

• Method 1 (HPLC): Instrument: HP 1100 with DAD detection; Pillar: Kromasil RP-18, 60 mm × 2.1 mm, 3.5 μm; Eluent A: 5 ml perchloric acid / l Water, eluent B: acetonitrile; Gradient: 0 min 2% B, 0.5 min 2% B, 4.5 minutes 90% B, 9 minutes 90% B, 9.2 minutes 2% B, 10 minutes 2% B; Flow: 0.75 ml / min; Oven: 30 ° C; UV detection: 210 nm.
• Method 2 (GC-MS): Instrument: Micromass GCT, GC6890; Column: Restek RTX-35MS, 30 m × 250 μm × 0.25 μm; constant Flow with helium: 0.88 ml / min; Oven: 60 ° C; Inlet: 250 ° C; Gradient: 60 ° C (0.30 min.), 50 ° C / min → 120 ° C, 16 ° C / min → 250 ° C, 30 ° C / min → 300 ° C (1.7 min hold).
• Method 3 (HPLC): Instrument: HP 1100 with DAD detection; Column: Kromasil RP-18, 60 mm × 2.1 mm, 3.5 μm; Eluent A: 5 ml perchloric acid / l Water, eluent B: acetonitrile; Gradient: 0 min 2% B, 0.5 min 2% B, 4.5 minutes 90% B, 6.5 minutes 90% B, 6.7 minutes 2% B, 7.5 minutes 2% B; River: 0.75 ml / min; Oven: 30 ° C; UV detection: 210 nm.
• Method 4 (LC-MS): Device Type MS: Micromass ZQ; device type HPLC: Waters Alliance 2795; Pillar: Phenomenex Synergi 2μ Hydro-RP Mercury 20mm × 4mm; Eluent A: 1 l of water + 0.5 ml of 50% formic acid, eluent B: 1 l of acetonitrile + 0.5 ml of 50% formic acid; Gradient: 0.0 min 90% A → 2.5 min 30% A → 3.0 min 5% A → 4.5 min 5% A; Flow: 0.0 min 1 ml / min, 2.5 min / 3.0 min / 4.5 min 2 ml / min; Oven: 50 ° C; UV detection: 210 nm.
• Method 5 (LC-MS): Device Type MS: Micromass ZQ; Device type HPLC: HP 1100 Series; W DAD; Column: Phenomenex Synergi 2μ Hydro-RP Mercury 20 mm × 4 mm; Eluent A: 1 l of water + 0.5 ml of 50% formic acid, eluent B: 1 l of acetonitrile + 0.5 ml of 50% formic acid; Gradient: 0.0 min 90% A → 2.5 min 30% A → 3.0 min 5% A → 4.5 min 5% A; Flow: 0.0 min 1 ml / min, 2.5 min / 3.0 min / 4.5 min. 2 ml / min; Oven: 50 ° C; UV detection: 210 nm.
• Method 6 (LC-MS): Instrument: Micromass Quattro LCZ with HPLC Agilent 1100 series; Pillar: Phenomenex Synergi 2μ Hydro-RP Mercury 20 mm × 4 mm; Eluent A: 1 l of water + 0.5 ml of 50% formic acid, eluent B: 1 liter acetonitrile + 0.5 ml 50% formic acid; Gradient: 0.0 min 90% A → 2.5 min 30% A → 3.0 min 5% A → 4.5 min 5% A; Flow: 0.0 min 1 ml / min, 2.5 min / 3.0 min / 4.5 min 2 ml / min; Oven: 50 ° C; UV detection: 208-400 nm.
• Method 7 (LC-MS): Instrument: Micromass Platform LCZ with HPLC Agilent 1100 series; Pillar: Thermo HyPURITY Aquastar 3μ 50 mm × 2.1 mm; Eluent A: 1 l of water + 0.5 ml of 50% formic acid, eluent B: 1 liter acetonitrile + 0.5 ml 50% formic acid; Gradient: 0.0 min 100% A → 0.2 min 100% A → 2.9 min 30% A → 3.1 min 10% A → 5.5 min 10% A; Oven: 50 ° C; Flow: 0.8 ml / min; UV detection: 210 nm.
• Method 8 (HPLC with chiral phase): Column: packing material Chiral Silica gel selector KBD 5287 (250 mm × 4.6 mm) based on the Selector poly (N-methacryloyl-L-leucine-dicyclopropylmethylamide); Eluent: methyl tert-butyl ether; Temperature: 24 ° C; River: 1 ml / min.
• Method 9 (HPLC with chiral phase): Column: packing material Chiral Silica gel selector KBD 5326 (600 mm × 40 mm) based on the Selector poly (N-methacryloyl-L-leucine-dicyclopropylmethylamide); Eluent A: Methyl tert-butyl ether, eluent B: tetrahydrofuran; Gradient: 0.0 min 100% A → 45 min 100% A → 55 min 100% B; Temperature: 24 ° C; River: 50 ml / min.
• Method 10 (LC-MS): Device Type MS: Micromass ZQ; device type HPLC: Waters Alliance 2790; Pillar: Grom-Sil 120 ODS-4 HE 50 mm × 2 mm, 3.0 μm; Eluent A: water + 500 μl 50% formic acid / l, Eluent B: acetonitrile + 500 μl 50% formic acid / L; Gradient: 0.0 min 70% B → 4.5 min 90% B; Oven: 45 ° C; Flow: 0.8 ml / min; UV detection: 210 nm.
• preparative RP-HPLC: column: YMC Gel ODS-AQ S-5/15 μM, 250 mm × 30 mm, eluent A: water, eluent B: acetonitrile; Gradient: 0.00 min 5% B → 3.00 min 5% B → 35.0 min 95% B → 46.0 min 95% B; Temp .: room temperature; Flow: 50 ml / min; UV detection.

Chemical synthesis of Examples

Synthesis of the starting compounds:

Synthesis of substituted beta-hydroxyphenylalanine derivatives by the example of methyl (betaR) -2- (benzyloxy) -5-bromo-N- (tert-butoxycarbonyl) -beta - {[tert-butyl (dimethyl) silyl] oxy} -L- phenylalaninate

Synthesis of protected biphenylbisamino acids exemplified by methyl (2S, 3R) -3- (4,4'-bis (benzyloxy) -3 '- {(2S) -2 - {[(benzyloxy) carbonyl] amino} - 3-oxo-3- [2- (trimethylsilyl) ethoxy] propyl} biphenyl-3-yl) -2 - [(tert-butoxycarbonyl) amino] -3 - {[tert-butyl (dimethyl) silyl] oxy} propane

starting compounds

Example 1A

Benzyl-2- (benzyloxy) -5-bromobenzoate

To a solution of 40 g (0.184 mol) of 5-bromo-2-hydroxybenzoic acid in 500 ml of anhydrous dimethylformamide are added 62 ml (0.46 mol) of benzylbromide and 76 g (0.553 mol) of potassium carbonate. After stirring for 16 h at 85 ° C, the cooled mixture is filtered and the filtrate is concentrated to dryness in vacuo. The residue is taken up in ethyl acetate and washed several times with water. The organic phase is dried over sodium sulfate, the solvent is evaporated in vacuo and the residue recrystallized from petroleum ether.
Yield: 58 g (79% of th.), Solid
HPLC (Method 1): R _t = 5.81 min.
MS (DCI (NH ₃ )): m / z = 414 (M + NH ₄ ) ⁺ .
¹ H-NMR (300 MHz, d ₆ -DMSO): δ = 5.2 (s, 2H), 5.3 (s, 2H), 7.23 (d, 1H), 7.25-7.45 (m, 10H) 7.70 (dd, 1H ), 7.8 (d, 1H).

Example 2A

2- (benzyloxy) -5-bromobenzoic

Representation see Collect. Czech. Chem. Commun., 1976, 41, 3384-3387. 30.6 g (77 mmol) of benzyl 2- (benzyloxy) -5-bromobenzoate (Example 1A) are dissolved in 450 ml of THF / water (1: 1) and treated with 77 ml of 1N sodium hydroxide solution. The reaction mixture is stirred at 80 ° C for 24 h. The mixture is concentrated in vacuo to a volume of 100 ml and extracted with diethyl ether. The aqueous phase is separated, adjusted to pH 1 with 1N hydrochloric acid and extracted several times with ethyl acetate. The organic phase is dried over magnesium sulfate and concentrated to dryness in vacuo.
Yield: 21 g (89% of theory), solid
GC-MS (Method 2): R _t = 12.90 min,
MS (EI +): m / z = 306 (M) ⁺ .
¹ H-NMR (400 MHz, d ₆ -DMSO): δ = 5.21 (s, 2H), 7.17 (d, 1H), 7.31 (m _c , 1H), 7.38 (m _c , 2H), 7.47 (m _c , 2H), 7.64 (dd, 1H), 7.73 (d, 1H), 13.05 (br. S, 1H).

Example 3A

Methyl-2- (benzyloxy) -5-bromo-N- (tert-butoxycarbonyl) -beta-oxophenylalaninat

18.8 ml (0.143 mmol) of 1-chloro-N, N under argon are added to a solution 1 of 17.5 g (57 mmol) of 2- (benzyloxy) -5-bromobenzoic acid (Example 2A) cooled to 0 ° C. in 80 ml of absolute dichloromethane , 2-trimethylpropenylamine added. It is stirred for 1 h at 0 ° C. 26.6 g (0.114 mmol) of methyl N- (tert-butoxycarbonyl) -3-oxoserate (see Synthesis, 1992, 1248-1254) in 770 ml of absolute THF at -70 ° C. with 143 ml are used under argon in a second flask (228 mmol) of a 1.6M solution of. Added n-butyllithium in hexane and stirred for 20 min. Subsequently, at -70 ° C, the acid chloride generated in solution 1 is slowly added dropwise. When the addition is complete, the mixture is warmed slowly to RT and stirred vigorously for a further 24 h. The solvents are evaporated in vacuo, the residue is taken up in ethyl acetate and washed successively with ice-cooled 0.5% sulfuric acid and 1N potassium bicarbonate solution. The organic phase is dried over magnesium sulfate and the solvent is concentrated in vacuo. The residue is purified by column chromatography on silica gel 60 (cyclohexane: ethyl acetate 20: 1).
Yield: 19 g (70% of theory), solid
HPLC (Method 3): R _t = 5.26 min.
MS (DCI (NH ₃ )): m / z = 495 (M + NH ₄ ) ⁺ .
¹ H-NMR (400 MHz, d ₆ -DMSO): δ = 1.35 (s, 9H), 3.54 (s, 3H), 5.25 (s, 2H), 5.68 (d, 1H), 7.2 (d, 1H) , 7.3-7.5 (m, 5H), 7.65-7.78 (m, 3H).

Example 4A

Methyl-2- (benzyloxy) -5-bromo-N- (tert-butoxycarbonyl) -beta-hydroxyphenylalaninat

2.1 ml of a 1M solution of (S) -2-methyl-CBS-oxazaborolidine in 25 ml of tetrahydrofuran are placed under argon and admixed with 4.8 ml (27 mmol) of borane-N, N-diethylaniline complex. After stirring at RT for 30 min, 10 g (20.9 mmol) of methyl 2- (benzyloxy) -5-bromo-N- (tert-butoxycarbonyl) -beta-oxophenylalaninate (Example 3A) dissolved in 25 ml of THF are added dropwise at 0 ° C. , After completion of the addition, the solution is stirred with slow warming to RT for 16 h. Then added dropwise 15 ml of methanol slowly and concentrated in vacuo. The residue is purified by chromatography on a preparative RP-HPLC. The product is available as a threo / erythro mixture (1: 5). The enantiomeric excess (ee) is in each case 55%, determined by analytical HPLC on a chiral phase (Method 8).
Yield: 5.1 g (51% of theory).
threo isomer:
LC-MS (Method 4): R _t = 2.59 min
MS (ESI): m / z = 480 (M + H) ⁺ .
¹ H-NMR (300 MHz, CDCl ₃ ): δ = 1.3 (s, 9H), 2.66 (br, s, 1H), 3.75 (s, 3H), 4.77 (br d, 1H), 5.14 (m _c , 2H), 5.25 (br s, 1H), 5.58 (br s, 1H), 6.8 (d, 1H), 7.28-7.50 (m, 6H), 7.55 (d, 1H).
erythro isomer:
LC-MS (Method 4): R _t = 2.57 min
MS (ESI): m / z = 480 (M + H) ⁺ .
¹ H-NMR (400 MHz, CDCl ₃ ): δ = 1.43 (s, 9H), 3.40 (s, 3H), 4.49 (br d, 1H), 4.71 (br, m, 1H), 5.05 (m _c , 2H), 5.34 (br, m, 1H), 5.40 (br, m, 1H), 6.77 (d, 1H), 7.28-7.45 (m, 6H), 7.47 (d, 1H).

Example 5A

Methyl (betar) -2- (benzyloxy) -5-bromo-N- (tert-butoxycarbonyl) -beta-hydroxy-L-phenylalaninate

The compound of Example 4A is separated as a mixture of isomers by chromatography on a chiral stationary phase (Method 9) in the four diastereomers. The target compound is eluted as the third fraction.
LC-MS (Method 6): R _t = 2.75 min
MS (ESI): m / z = 480 (M + H) ⁺ .

Example 6A

Methyl (betar) -2- (benzyloxy) -5-bromo-N- (tert-butoxycarbonyl) -beta - {[tert-butyl (dimethyl) silyl] oxy} -L-phenylalaninate

A solution of 200 mg (0.416 mmol) of methyl (betaR) -2- (benzyloxy) -5-bromo-N- (tert-butoxycarbonyl) -beta-hydroxy-L-phenylalaninate (Example 5A) in 2.5 ml of DMF is added 0 ° C with 220 mg (0.832 mmol) trifluoromethanesulfonic acid tert-butyldimethylsilylester, 0.084 mg (0.832 mmol) of triethylamine and 1 mg (0.42 mmol) of 4-dimethylaminopyridine and stirred after warming to RT for 24 hours. The reaction mixture is treated with dichloromethane and washed successively with 1N sodium bicarbonate solution and water. The combined organic phases are dried over magnesium sulfate, filtered, concentrated and dried in vacuo. The crude product is purified by chromatography (RP-HPLC, mobile phase gradient from acetonitrile / water).
Yield: 140 mg (57% of theory)
LC-MS (Method 5): R _t = 3.51 min
MS (ESI): m / z = 595 (M + H) ⁺ .
¹ H-NMR (300 MHz, CDCl ₃ ): δ = -0.15 (s, 3H), -0.7 (s, 3H), 0.87 (s, 9H), 1.29 (s, 9H), 3.72 (s, 3H) , 4.7 (dd, 1H), 5.0-5.3 (m, 3H), 5.68 (d, 1H), 6.76 (d, 1H), 7.25-7.50 (m, 7H).

Example 7A

2- (trimethylsilyl) ethyl-2- (benzyloxy) -N - [(benzyloxy) carbonyl] -5- (4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl) -L- phenylalaninate

To a solution of 2.0 g (3.17 mmol) of 2 (S) -benzyloxycarbonylamino-3- (2-benzyloxy-5-iodo-phenyl) -propionic acid (2-trimethylsilyl) -ethyl ester (Example 11A from WO03 / 106480) in 30 ml of DMSO are added 0.923 g (9.5 mmol) of potassium acetate. The mixture is deoxygenated by passing argon through the vigorously stirred solution for 15 minutes. Then, 0.924 g (3.64 mmol) of 4,4,4 ', 4', 5,5,5 ', 5'-octamethyl-2,2'-bi-1,3,2-dioxaborolane and 0.116 g (0.160 mmol , 0.05 equivalent) of bis (diphenylphosphino) ferrocenepalladium (II) chloride. Under gentle argon flow is heated to 80 ° C and cooled after 6 h again. The mixture is filtered through silica gel (eluent: dichloromethane). The residue is purified by column chromatography on silica gel (mobile phase: cyclohexane: ethyl acetate 4: 1).
Yield: 1.12 g (56% of theory)
LC-MS (Method 10): R _t = 3.02 min.
MS (EI): m / z = 632 (M + H) ^+
1 H-NMR (200 MHz, CDCl _3): δ = 0.92 (dd, 2H), 1.31 (s, 12H), 2.95-3.95 (m, 2H), 4.11 (m _c, 2H), 4:55 (11 (m _c , 1H), 4.99 (s, 2H), 5.08 (s, 2H), 5.53 (d, 1H), 6.90 (d, 1H), 7.15-7.47 (m, 10H), 7.58 (d, 1H), 7.67 (dd, 1H).

Example 8A

Methyl (2S, 3R) -3- (4,4-bis (benzyloxy) -3 '- {(2S) -2 - {[(benzyloxy) carbonyl] amino} -3-oxo-3- [2- (trimethylsilyl) ethoxy] propyl} biphenyl-3-yl) -2 - [(tert-butoxycarbonyl) amino] -3 - {[tert-butyl (dimethyl) silyl] oxy} propanoate

A solution of 2.0 g (3.36 mmol) of methyl (betaR) -2- (benzyloxy) -5-bromo-N- (tert-butoxycarbonyl) -beta - {[tert-butyl (dimethyl) silyl] oxy} - L-phenylalaninate (Example 6A) and 2.92 g (3.70 mmol) of 2- (trimethylsilyl) ethyl 2- (benzyloxy) -N - [(benzyloxy) carbonyl] -5- (4,4,5,5-tetramethyl-1 , 3,2-dioxaborolan-2-yl) -L-phenylalaninate (Example 7A) in 16 ml of 1-methyl-2-pyridone is rendered inert and saturated with argon (about 30 minutes to pass argon). Subsequently, 250 mg (0.34 mmol) of bis (diphenylphosphino) ferrocene-palladium (II) chloride (PdCl ₂ (dppf)), 2.19 g (6.73 mmol) of cesium carbonate and 0.8 ml of water are added. The reaction mixture is poured over with argon and stirred for 3 h at 50 ° C. The mixture is cooled, taken up in ethyl acetate and washed successively with saturated sodium chloride solution and water. The organic phase is dried over magnesium sulfate and the solvent is concentrated in vacuo. The residue is purified by column chromatography on silica gel (cyclohexane: ethyl acetate gradient: 15: 1 → 10: 1) and then stirred in methanol. The resulting solid is filtered off and dried under high vacuum.
Yield: 2.77 g (77% of theory).
LC-MS (Method 5): R _t = 3.75 min
MS (EI): m / z = 1019 (M + H) ⁺ .
¹ H-NMR (300 MHz, CDCl _3): δ = -0.11 (s, 3H), 0.00 (m, 12H), 0.8-0.95 (m, 11H), 1.17 (s, 9H), 3.0-3.3 (m , 2H), 3.75 (s, 3H), 4.0-4.25 (m, 2H), 4.52-4.65 (m, 1H), 4.77 (m _c, 1H), 4.92-5.3 (m, 7H), 5.52-5.65 ( m, 1H), 5.77 (d, 1H), 6.85-6.97 (m, 2H), 7.19-7.46 (m, 16H), 7.48-7.65 (m, 3H).

Example 9A

Methyl (2S, 3R) -2-amino-3- (4,4-bis (benzyloxy) -3 '- {(2S) -2 - {[(benzyloxy) carbonyl] amino} -3-oxo-3 - [2- (trimethylsilyl) ethoxy] propyl} biphenyl-3-yl) -3 - {[tert-butyl (dimethyl) silyl] oxy} propanoate hydrochloride

To a cooled to 0 ° C solution of 1.64 g (1.61 mmol) of methyl (2S, 3R) -3- (4,4'-bis (benzyloxy) -3 '- {(2S) -2 - {[(benzyloxy ) carbonyl] amino} -3-oxo-3- [2- (trimethylsilyl) ethoxy] propyl} biphenyl-3-yl) -2 - [(tert-butoxycarbonyl) amino] -3 - {[tert-butyl (dimethyl ) Silyl] oxy} propanoate (Example 8A) in 5 ml of anhydrous dioxane are added 30 ml of a 4M hydrogen chloride-dioxane solution over about 20 minutes. After stirring for 3 h, the solvent is evaporated in vacuo, the crude product dried under high vacuum and used without further purification for subsequent syntheses.
Yield: quant.
LC-MS (Method 4): R _t = 3.24 min.
MS (EI): m / z = 919 (M-HCl + H) ⁺ .

Example 10A

Methyl (2S, 3R) -2 - [((2S, 4R) -5 - {[(benzyloxy) carbonyl] amino} -2 - [(tert-butoxycarbonyl) amino] -4 - {[tert-butyl (dimethyl ) silyl] oxy} pentanoyl) amino] -3- (4,4-bis (benzyloxy) -3 '- {(2S) -2 - {[(benzyloxy) carbonyl] amino} -3-oxo-3 - [2- (trimethylsilyl) ethoxy] propyl} biphenyl-3-yl) -3 - {[tert-butyl (dimethyl) silyl] oxy} propanoate

To a cooled to 0 ° C solution of 1.54 g (1.61 mmol) of methyl (2S, 3R) -2-amino-3- (4,4'-bis (benzyloxy) -3 '- {(2S) -2- {[(benzyloxy) carbonyl] amino} -3-oxo-3- [2- (trimethylsilyl) ethoxy] -propyl} biphenyl-3-yl) -3 - {[tert-butyl (dimethyl) silyl] oxy} propanoate hydrochloride (Example 9A) and 0.88 g (1.77 mmol) of 5-benzyloxycarbonylamino-2 (S) -tert-butoxycarbonylamino-4 (R) - (tert-butyldimethylsilyloxy) -pentanoic acid (Example 14A from WO03 / 106480) in 25 ml of anhydrous DMF 0.76 g (2.01 mmol) of HATU and 0.58 g (4.5 mmol) of N, N-diisopropylethylamine were added. After stirring for 30 minutes at 0 ° C., an additional 0.15 g (1.13 mmol) of N, N-diisopropylethylamine are added. The reaction mixture is stirred at RT for 15 h. The solvent is then evaporated and the residue taken up in dichloromethane. The organic phase is washed successively with water, 0.2N hydrochloric acid and saturated aqueous sodium chloride washed solution, dried over sodium sulfate and concentrated. The crude product is purified by column chromatography on silica gel with dichloromethane / ethyl acetate (gradient 30/1 → 20/1 → 2/1).
Yield: 1.94 g (86% of theory)
LC-MS (Method 4): R _t = 3.98 min.
MS (EI): m / z = 1397 (M + H) ⁺ .

Example 11A

Methyl (2S, 3R) -2 - ({(2S) -3 - {[(benzyloxy) carbonyl] amino} -2 - [(tert-butoxycarbonyl) amino] butanoyl} amino) -3- (4,4 '-bis (benzyloxy) -3' - {(2S) -2 - {[(benzyloxy) carbonyl] amino} -3-oxo-3- [2- (trimethylsilyl) ethoxy] propyl} biphenyl-3-yl) - 3 - {[tert-butyl (dimethyl) silyl] oxy} propanoate

The preparation is carried out analogously to Example 10A from 0.352 g (0.37 mmol) of methyl (2S, 3R) -2-amino-3- (4,4'-bis (benzyloxy) -3 '- {(2S) -2 - [ (benzyloxy) carbonyl] amino} -3-oxo-3- [2- (trimethylsilyl) ethoxy] -propyl} biphenyl-3-yl) -3 - {[tertbutyl (dimethyl) silyl] oxy} propanoate hydrochloride (Example 9A) and 142 mg (0.41 mmol) of (2S) -4 - {[(benzyloxy) carbonyl] amino} -2 - [(tert-butoxycarbonyl) amino] butanecarboxylic acid with 154 mg (0.41 mmol) HATU and a total of 167 mg ( 1.29 mmol) Hünig's base in 5 ml of absolute DMF.
Yield: 412 mg (72% of theory)
LC-MS (Method 4): R _t = 3.55 min.
MS (EI): m / z = 1253 (M + H) ⁺ .

Example 12A

Methyl (2S, 3R) -2 - ({(2S) -5 - {[(benzyloxy) carbonyl] amino} -2 - [(tert-butoxycarbonyl) amino] pentanoyl} amino) -3- (4,4 '-bis (benzyloxy) -3' - {(2S) -2 - {[(benzyloxy) carbonyl] amino} -3-oxo-3- [2- (trimethylsilyl) ethoxy] propyl} biphenyl-3-yl) - 3 - {[tert-butyl (dimethyl) silyl] oxy} propanoate

The preparation is analogous to Example 10A from 0.98 g (1.02 mmol) of methyl (2S, 3R) -2-amino-3- (4,4'-bis (benzyloxy) -3 '- {(2S) -2 - {[ (benzyloxy) carbonyl] amino} -3-oxo-3- [2- (trimethylsilyl) ethoxy] -propyl} biphenyl-3-yl) -3 - {[tertbutyl (dimethyl) silyl] oxy} propanoate hydrochloride (Example 9A) and 0.45 g (1.23 mmol) of N ⁵ - [(benzyloxy) carbonyl] -N ² - (tert-butoxycarbonyl) -L-ornithine with 0.486 g (1.28 mmol) of HATU and a total of 0.463 g (3.58 mmol) Hünig's base in 16 ml of absolute DMF.
Yield: 1.06 g (68% of theory)
LC-MS (Method 4): R _t = 3.56 min.
MS (EI): m / z = 1267 (M + H) ⁺ .

Example 13A

Methyl (2S, 3R) -2 - ({(2S) -3 - {[(benzyloxy) carbonyl] amino} -2 - [(tert-butoxycarbonyl) amino] hexanoyl} amino) -3- (4,4 '-bis (benzyloxy) -3' - {(2S) -2 - {[(benzyloxy) carbonyl] amino} -3-oxo-3- [2- (trimethylsilyl) ethoxy] propyl} biphenyl-3-yl) - 3 - {[tert-butyl (dimethyl) silyl] oxy} propanoate

The preparation is carried out analogously to Example 10A from 0.38 g (0.40 mmol) of methyl (2S, 3R) -2-amino-3- (4,4'-bis (benzyloxy) -3 '- {(2S) -2 - [ (benzyloxy) carbonyl] amino} -3-oxo-3- [2- (trimethylsilyl) ethoxy] -propyl} biphenyl-3-yl) -3 - {[tertbutyl (dimethyl) silyl] oxy} propanoate hydrochloride (Example 9A) and 0.18 g (0.48 mmol) of N ⁶ - [(benzyloxy) carbonyl] -N ² - (tert-butoxycarbonyl) -L-lysine with 0.188 g (0.50 mmol) of HATU and a total of 0.180 g (1.39 mmol) of Hünig base in 5 ml of absolute DMF.
Yield: 0.63 g (78% of theory)
LC-MS (Method 4): R _t = 3.57 min.
MS (EI): m / z = 1281 (M + H) ⁺ .

Example 14A

(2S) -2 - {[(benzyloxy) carbonyl] amino} -3- {4,4-bis (benzyloxy) -3 '- [(1R, 2S) -2 - ({(2S, 4R) -5 - {[(benzyloxy) carbonyl] amino} -2 - [(tert-butoxycarbonyl) amino] -4-hydroxypentanoyl} amino) -1-hydroxy-3-methoxy-3-oxopropyl] biphenyl-3-yl} -propionic acid

To a solution of 183 mg (0.13 mmol) of methyl (2S, 3R) -2 - [((2S, 4R) -5 - {[(benzyloxy) carbonyl] amino} -2 - [(tert-butoxycarbonyl) amino ] -4 - {[tert-butyl (dimethyl) silyl] oxy} pentanoyl) amino] -3- (4,4-bis (benzyloxy) -3 '- {(2S) -2 - {[(benzyloxy) carbonyl ] amino} -3-oxo-3- [2- (tri methylsilyl) ethoxy] -propyl} biphenyl-3-yl) -3 - {[tert-butyl (dimethyl) silyl] oxy} propanoate (Example 10A) in 2.65 ml of absolute DMF at RT is added dropwise 0.52 ml of 1N tetrabutylammonium fluoride solution in THF added. After 60 min at RT is cooled to 0 ° C and treated with ice water. It is immediately mixed with ethyl acetate and a little 1N hydrochloric acid solution. The organic phase is dried over magnesium sulfate, concentrated in vacuo and dried for 1 h under high vacuum. The crude product is reacted without further purification.
Yield: quant.
LC-MS (Method 4): R _t = 2.82 min.
MS (EI): m / z = 1069 (M + H) ⁺ .

Analogous The procedure of Example 14A is set forth in the following table listed Examples 15A to 17A made.

Example 18A

Methyl (2S, 3R) -2 - ({(2S, 4R) -5 - {[(benzyloxy) carbonyl] amino} -2 - [(tert-butoxycarbonyl) amino] -4-hydroxypentanoyl} amino) -3- {4,4'-bis (benzyloxy) -3 '- [(2S) -2 - {[(benzyloxy) carbonyl] amino} -3-oxo-3- (pentafluorophenoxy) propyl] biphenyl-3-yl} -3 -hydroxypropanoat

141 mg (crude product, ca. 0.13 mmol) of (2S) -2 - {[(benzyloxy) carbonyl] amino} -3- {4,4'-bis (benzyloxy) -3 '- [(1R, 2S) - 2 - ({(2S, 4R) -5 - {[(benzyloxy) carbonyl] amino} -2 - [(tert-butoxycarbonyl) amino] -4-hydroxypentanoyl} amino) -1-hydroxy-3-methoxy-3 -oxopropyl] biphenyl-3-yl} -propionic acid (Example 14A) are initially charged in 5 ml of dichloromethane and treated with 121 mg (0.66 mmol) of pentafluorophenol, dissolved in 1 ml of dichloromethane. Add 2 mg (0.01 mmol) of DMAP and cool to -25 ° C (ethanol / carbon dioxide bath). At -25 ° C, 39 mg (0.172 mmol) of EDC are added. The mixture warms slowly overnight to RT. The reaction mixture is concentrated in vacuo and dried briefly in a high vacuum. The crude product is reacted without further purification.
Yield: quant.
LC-MS (method 5): R _t = 3.34 min.
MS (EI): m / z = 1235 (M + H) ⁺ .

Analogous for the prescription of Example 18A are those in the following table listed Examples 19A to 21A made.

Example 22A

Methyl (2S, 3R) -2 - [((2S, 4R) -2-amino-5 - {[(benzyloxy) carbonyl] amino} -4-hydroxypentanoyl) amino] -3- {4,4 bis (benzyloxy) -3 '- [(2S) -2 - {[(benzyloxy) carbonyl] amino} -3-oxo-3- (pentafluorophenoxy) propyl] biphenyl-3-yl} -3-hydroxypropanoate hydrochloride

0.163 g (crude, ca. 0.132 mmol) of methyl (2S, 3R) -2 - ({(2S, 4R) -5 - {[(benzyloxy) carbonyl] amino} -2 - [(tert-butoxycarbonyl) amino ] -4-hydroxypentanoyl} amino) -3- {4,4-bis (benzyloxy) -3 '- [(2S) -2 - {[(benzyloxy) carbonyl] amino} -3-oxo-3- (pentafluorophenoxy ) propyl] biphenyl-3-yl} -3-hydroxypropanoate (Example 18A) are dissolved at 0 ° C in 4 ml of a 4M hydrogen chloride-dioxane solution. After 2.5 h at 0 ° C, the reaction solution is concentrated in vacuo, coevaporated several times with dichloromethane and dried under high vacuum. The crude product is reacted without further purification.
Yield: quant.
LC-MS (method 5): R _t = 2.71 min.
MS (EI): m / z = 1135 (M-HCl + H) ⁺ .

Analogous to the example of Example 22A are those in the following table listed Examples 23A to 25A made.

Example 26A

Methyl (7R, 8S, 11S, 14S) -5.17-bis (benzyloxy) -14 - {[(benzyloxy) carbonyl] amino} -11 - ((2R) -3 - {[(benzyloxy) carbonyl] amino } -2-hydroxypropyl) -7-hydroxy-10,13-dioxo-9,12-diazatricyclo [14.3.1.1 ^2,6 ] henicosa-1 (20), 2 (21), 3,5,16,18- hexaene-8-carboxylate

150 mg (crude, ca. 0.13 mmol) of methyl (2S, 3R) -2 - [((2S, 4R) -2-amino-5 - {[(benzyloxy) carbonyl] amino} -4-hydroxypentanoyl) amino ] -3- {4,4-bis (benzyloxy) -3 '- [(2S) -2 - {[(benzyloxy) carbonyl] amino} -3-oxo-3- (pentafluorophenoxy) propyl] biphenyl-3 -yl} -3-hydroxypropanoate hydrochloride are dissolved in 150 ml of dichloromethane and treated dropwise with a solution of 0.37 ml of triethylamine in 30 ml of dichloromethane. It is stirred overnight at RT. For workup, the mixture is gently evaporated in vacuo and the residue is stirred in acetonitrile. The solid is filtered off, washed with acetonitrile and dried under high vacuum.
Yield: 73 mg (58% of theory, over four steps starting from Example 10A).
LC-MS (method 5): R _t = 3.02 min.
MS (EI): m / z = 951 (M + H) ⁺ .
¹ H NMR (300 MHz, d ₆ -DMSO): δ = 1.42-1.61 (m, 1H), 1.7-1.95 (m, 1H), 2.92 (m _c , ¹ H), 3.02-3.13 (m, 2H) , 3.3 (m, 1H under D ₂ O), 3.68 (s, 3H), 4.45 (m _c , 1H), 4.68 (m _c , 1H), 4.83-5.25 (m, 9H), 5.77 (d, 1H) , 5.86 (d, 1H), 6.53 (d, 1H), 6.95-7.2 (m, 4H), 7.21-7.60 (m, 23H), 8.54 (d, 1H), 8.64 (d, 1H).

Analogous The procedure of Example 26A is set forth in the following table listed Examples 27A to 29A made.

Example 30A

(7R, 8S, 11S, 14S) -5.17-bis (benzyloxy) -14 - {[(benzyloxy) carbonyl] amino} -11 - ((2R) -3 - {[(benzyloxy) carbonyl] amino} - 2-hydroxypropyl) -7-hydroxy-10,13-dioxo-9,12-diazatricyclo [14.3.1.1 ^2,6 ] -henicosa-1 (20), 2 (21), 3,5,16,18-hexaene -8-carboxylic acid

A suspension of 25 mg (0.026 mmol) of methyl (7R, 8S, 11S, 14S) -5,17-bis (benzyloxy) -14 - {[(benzyloxy) carbonyl] amino} -11 - ((2R) -3 - {[(benzyloxy) carbonyl] amino} -2-hydroxypropyl) -7-hydroxy-10,13-dioxo-9,12-diazatricyclo [14.3.1.1 ^2,6 ] henicosa-1 (20), 2 (21) , 3,5,16,18-Hexaene-8-carboxylate (Example 26A) in 10 ml dioxane / water (4: 1) is added after addition of 0.53 ml (0.053 mmol) of a 0.1N aqueous lithium hydroxide solution for 3 h 10 ° C stirred. By addition of 0.1N hydrochloric acid is adjusted to pH 2, the precipitated solid is filtered off and dried under high vacuum.
Yield: 24 mg (98% of theory)
LC-MS (Method 6): R _t = 2.85 min.
MS (EI): m / z = 937 (M + H) ⁺ .

Example 31A

(7R, 8S, 11S, 14S) -5.17-bis (benzyloxy) -14 - {[(benzyloxy) carbonyl] amino} -11- (3 - {[(benzyloxy) carbonyl] amino} propyl) -7 -hydroxy-10,13-dioxo-9,12-diazatricyclo [14.3.1.1 ^2,6 ] henicosa-1 (20), 2 (21), 3,5,16,18-hexaene-8-carboxylic acid

A suspension of 30 mg (0.032 mmol) of methyl (7R, 8S, 11S, 14S) -5,17-bis (benzyloxy) -14 - {[(benzyloxy) carbonyl] amino} -11- (3 - {[( benzyloxy) carbonyl] amino} propyl) -7-hydroxy-10,13-dioxo-9,12-diazatricyclo [14.3.1.1 ^2,6 ] henicosa-1 (20), 2 (21), 3,5,16, 18-Hexaene-8-carboxylate (Example 28A) in 25 ml of tetrahydrofuran / water (4: 1) is stirred for 12 h at RT after addition of 0.64 ml (0.054 mmol) of a 0.1N aqueous lithium hydroxide solution. By addition of 0.1N hydrochloric acid is adjusted to pH 2, the precipitated solid is filtered off and dried under high vacuum.
Yield: 25 mg (85% of theory)
LC-MS (method 5): R _t = 2.95 min.
MS (EI): m / z = 921 (M + H) ⁺ .

Analogous The procedure of Example 31A is that given in the following table listed Examples 32A and 33A made.

Example 34A

Benzyl - {(2R) -3 - [(7R, 8S, 11S, 14S) -8 - {[(2-aminoethyl) amino] carbonyl} -5.17-bis (benzyloxy) -14 - {[(benzyloxy) carbonyl] amino} -7-hydroxy-10,13-dioxo-9,12-diazatricyclo [14.3.1.1 ^2,6 ] henicosa-1 (20), 2 (21), 3,5,16,18-hexaene 11-yl] -2-hydroxypropyl} carbamate

A solution of 40 mg (0.042 mmol) of methyl (7R, 8S, 11S, 14S) -5,17-bis (benzyloxy) -14 - {[(benzyloxy) carbonyl] amino} -11 - ((2R) -3 - {[(benzyloxy) carbonyl] amino} -2-hydroxypropyl) -7-hydroxy-10,13-dioxo-9,12-diazatricyclo [14.3.1.1 ^2,6 ] henicosa-1 (20), 2 (21) , 3,5,16,18-hexaene-8-carboxylate (Example 26A) in 1.5 ml of 1,2-diaminoethane is stirred at RT after addition of 0.55 mg (0.008 mmol) of potassium cyanide for 16 h. After addition of water, the precipitated solid is filtered off and dried under high vacuum.
Yield: 27 mg (65% of theory)
LC-MS (Method 5): R _t = 2.39 min.
MS (EI): m / z = 979 (M + H) ⁺ .

Analogous The procedure of Example 34A is set forth in the following table listed Examples 35A to 37A made.

Example 38A

Benzyl- {2 - [(7R, 8S, 11S, 14S) -5.17-bis (benzyloxy) -14 - {[(benzyloxy) carbonyl] amino} -8 - [({2- [2-aminoethyl-bis ( ) amino] ethyl} amino) carbonyl] -7-hydroxy-10,13-dioxo-9,12-diazatricyclo [14.3.1.1 ^2,6 ] -henicosa-1 (20), 2 (21), 3,5, 16,18-hexaene-11-yl] ethyl} carbamate

The preparation is carried out analogously to Example 34A from 15 mg (0.02 mmol) Methyl (7R, 8S, 11S, 14S) -5.17-bis (benzyloxy) -14 - {[(benzyloxy) carbonyl] amino} -11- (2 - {[(benzyloxy) carbonyl] amino} ethyl) 7-hydroxy-10,13-dioxo-9,12-diazatricyclo [14.3.1.1 ^2,6 ] henicosa-1 (20), 2 (21), 3,5,16,18-hexaene-8-carboxylate ( Example 27A) in 0.5 ml of N, N-bis (2-aminoethyl) ethane-1,2-diamine and with 0.11 mg (0.0016 mmol) of potassium cyanide.
Yield: 14.4 mg (85% of theory)
LC-MS (Method 5): R _t = 1.99 min.
MS (EI): m / z = 1035 (M + H) ⁺ .

Example 39A

Methyl (7R, 8S, 11S, 14S) -14-amino-11 - [(2R) -3-amino-2-hydroxypropyl] -5,7,17-trihydroxy-10,13-dioxo-9,12- diazatricyclo [14.3.1.1 ^2,6 ] henicosa-1 (20), 2 (21), 3,5,16,18-hexaene-8-carboxylate dihydrochloride

There are 30 mg (0.32 mmol) of methyl (7R, 8S, 11S, 14S) -5,17-bis (benzyloxy) -14 - {[(benzyloxy) carbonyl] amino} - ((2R) -3- { [(benzyloxy) carbonyl] amino} -2-hydroxypropyl) -7-hydroxy-10,13-dioxo-9,12-diazatricyclo [14.3.1.1 ^2,6 ] henicosa-1 (20), 2 (21), 3 , 5,16,18-Hexaene-8-carboxylate (Example 26A) in a mixture of 10 ml of acetic acid / water / ethanol 4: 1: 1. 10 mg of palladium on activated carbon (10% strength) are added thereto and the mixture is subsequently hydrogenated at normal pressure for 15 hours. The reaction mixture is filtered through prewashed diatomaceous earth and the filtrate evaporated in vacuo. The residue is treated with 0.95 ml of 0.1N aqueous hydrochloric acid and concentrated. The residue is stirred with 10 ml of diethyl ether and decanted off. The remaining solid is dried under high vacuum.
Yield: 18 mg (97% of theory).
LC-MS (Method 7): R _t = 2.29 min
MS (EI): m / z = 503 (M-2HCl + H) ⁺ .
¹ H-NMR (400 MHz, D ₂ O): δ = 1.8-1.93 (m, 1H), 1.96-2.1 (m, 1H), 2.83-2.93 (m, 1H), 2.97-3.12 (m, 2H) , 3.5 (m _c , 1H), 3.77 (s, 3H), 3.95-4.05 (m, 1H), 4.37-4.42 (m, 1H), 4.9 (m _c , 1H), 4.94-4.49 (m, 1H) , 5.78 (m _c, 1H), 6.84-6.95 (m, 3H), 7:27 to 7:42 (m, 3H).

Analogous for the example of Example 39A are those in the following table listed Examples 40A-46A made.

Example 47A

(7R, 8S, 11S, 14S) -14 - [(tert-butoxycarbonyl) amino] -11 - {(2R) -3 - [(tert-butoxycarbonyl) amino] -2-hydroxypropyl} -5,7,17- trihydroxy-10,13-dioxo-9,12-diazatricyclo [14.3.1.1 ^2,6 ] henicosa-1 (20), 2 (21), 3,5,16,18-hexaene-8-carboxylic acid

50 mg (0.087 mmol) of methyl (7R, 8S, 11S, 14S) -14-amino-11 - [(2R) -3-amino-2-hydroxypropyl] -5,7,17-trihydroxy-10,13- dioxo-9,12-diazatricyclo [14.3.1.1 ^2,6 ] henicosa-1 (20), 2 (21), 3,5,16,18-hexaene-8-carboxylate dihydrochloride (Example 39A) are dissolved in 0.43 ml ( 0.43 mmol) dissolved 1N sodium hydroxide solution and at room temperature with stirring with 56.9 mg (0.26 mmol) of di-tert-butyl-dicarbonate, dissolved in 0.5 ml of methanol. After stirring for 2 h at RT is determined by Zutrop of 0.1N hydrochloric acid to pH 3. It is extracted three times with ethyl acetate, the organic phase is dried over magnesium sulfate and concentrated by evaporation in vacuo to constant weight.
Yield: 37 mg (62% of theory).
LC-MS (Method 6): R _t = 1.83 min.
MS (EI): m / z = 703 (M + H) ⁺ .

Example 48A

(7R, 8S, 11S, 14S) -14 - [(tert-butoxycarbonyl) amino] -11- {3 - [(tert-butoxycarbonyl) amino] propyl} -5,7,17-trihydroxy-10,13-dioxo -9,12-diazatricyclo [14.3.1.1 ^2,6 ] henicosa-1 (20), 2 (21), 3,5,16,18-hexaene-8-carboxylic acid

The preparation is carried out analogously to Example 47A from 50 mg (0.088 mmol) of methyl (7R, 8S, 11S, 14S) -14-amino-11- (3-aminopropyl) -5,7,17-trihydroxy-10,13-dioxo -9,12-diazatricyclo [14.3.1.1 ^2,6 ] -henicosa-1 (20), 2 (21), 3,5,16,18-hexaene-8-carboxylate dihydrochloride (Example 41A) with 58 mg (0.26 mmol) of di-tert-butyl-dicarbonate in 0.44 ml (0.44 mmol) of 1N sodium hydroxide solution and 0.5 ml of methanol.
Yield: 46 mg (77% of theory).
LC-MS (Method 6): R _t = 1.93 min.
MS (EI): m / z = 687 (M + H) ⁺ .

Example 49A

tert-Butyl - [(1S) -4 - [(tert-butoxycarbonyl) amino] -1- (hydroxymethyl) butyl] carbamate

A solution of 300 mg (0.90 mmol) of N ² , N ⁵ -bis (tert-butoxycarbonyl) -L-ornithine in 10 ml of tetrahydrofuran at -10 ° C with 91 mg (0.90 mmol) of 4-methylmorpholine and 98 mg (0.90 mmol) of ethyl chloroformate and stirred for 30 min. At this temperature, 1.81 ml (1.81 mmol) of a 1M solution of lithium aluminum hydride in tetrahydrofuran are slowly added dropwise. It is slowly warmed to RT and stirred for 12 h at RT. With ice-cooling, carefully add 0.1 ml of water and 0.15 ml of 4.5% sodium hydroxide solution and stir for a further 3 h at RT. The batch is filtered and the filtrate is concentrated in vacuo. The residue is dissolved in ethyl acetate, washed with water, dried over magnesium sulfate and again concentrated to dryness in vacuo. The product is reacted without further purification.
Yield: 239 mg (83% of theory)
MS (ESI): m / z = 319 (M + H) ⁺ ; 341 (M + Na) ⁺ .

Example 50A

(2S) -2,5-Bis [(tert-butoxycarbonyl) amino] pentyl methanesulfonate

A solution of 240 mg (0.75 mmol) tert-butyl - [(1S) -4 - [(tert-butoxycarbonyl) amino] -1- (hydroxymethyl) butyl] carbamate (Example 49A) in 20 ml dichloromethane is added with 103 mg ( 0.90 mmol) of methanesulfonyl chloride and 0.21 ml (1.5 mmol) of triethylamine and stirred for 16 h at RT. It is diluted with dichloromethane and washed twice with 0.1N hydrochloric acid. The organic phase is dried over magnesium sulfate and concentrated in vacuo to dryness. The product is reacted without further purification.
Yield: 218 mg (73% of theory)
MS (ESI): m / z = 419 (M + Na) ⁺ .

Example 51A

tert-Butyl - {(4S) -5-azido-4 - [(tert-butoxycarbonyl) amino] pentyl} carbamate

A solution of 218 mg (0.55 mmol) of (2S) -2,5-bis [(tert-butoxycarbonyl) amino] pentyl methanesulfonate (Example 50A) in 15 ml of dimethylformamide is combined with 36 mg (0.55 mmol) of sodium azide and stirred for 12 h stirred at 70 ° C. Most of the solvent is distilled off in vacuo and the residue is diluted with ethyl acetate. It is washed several times with saturated sodium bicarbonate solution, dried over magnesium sulfate and concentrated to dryness in vacuo. The product is reacted without further purification.
Yield: 188 mg (99% of theory)
MS (ESI): m / z = 344 (M + H) ⁺ .

Example 52A

tert-Butyl - {(4S) -5-amino-4 - [(tert-butoxycarbonyl) amino] pentyl} carbamate

A solution of 188 mg (0.55 mmol) of tert-butyl {(4S) -5-azido-4 - [(tert-butoxycarbonyl) amino] pentyl} carbamate (Example 51A) in ethanol is added after addition of 20 mg of palladium Activated carbon (10%) hydrogenated at RT and normal pressure for 12 h. It is filtered through kieselguhr and the residue is washed with ethanol. The filtrate is concentrated to dryness in vacuo. The product is reacted without further purification.
Yield: 102 mg (59% of theory)
MS (ESI): m / z = 318 (M + H) ⁺ ; 340 (M + Na) ⁺ .

Example 53A

tert-Butyl - {(3S) -3 - [(tert-butoxycarbonyl) amino] -4-hydroxybutyl} carbamate

The preparation is carried out analogously to Example 49A from 300 mg (0.60 mmol) of dicyclohexylamino (2S) -2,4-bis [(tert-butoxycarbonyl) amino] butanate in 10 ml of tetrahydrofuran with 61 mg (0.60 mmol) of 4-methylmorpholine, 65 mg (0.60 mmol) ethyl chloroformate and 1.2 ml (1.20 mmol) of a 1M solution of lithium aluminum hydride in tetrahydrofuran. The product is reacted without further purification.
Yield: 174 mg (95% of theory)
MS (ESI): m / z = 305 (M + H) ⁺ .

Example 54A

(2S) -2,4-Bis [(tert-butoxycarbonyl) amino] butyl methanesulfonate

The preparation is analogous to Example 50A from 250 mg (0.81 mmol) tert-butyl - {(3S) -3 - [(tert-butoxycarbonyl) amino] -4-hydroxybutyl} carbamate (Example 53A) in 20 ml of dichloromethane with 110 mg ( 0.97 mmol) of methanesulfonyl chloride and 0.23 ml (1.6 mmol) of triethylamine. The product is reacted without further purification.
Yield: 200 mg (64% of theory)
MS (ESI): m / z = 383 (M + H) ⁺ ; 400 (M + Na) ⁺ .

Example 55A

tert-Butyl - {(3S) -4-azido-3 - [(tert-butoxycarbonyl) amino] butyl} carbamate

The preparation is carried out analogously to Example 51A from 200 mg (0.52 mmol) of (2S) -2,4-bis [(tert-butoxycarbonyl) amino] butyl methanesulfonate (Example 54A) in 15 ml of dimethylformamide with 34 mg (0.52 mmol) of sodium azide. The product is reacted without further purification.
Yield: 171 mg (99% of theory)

Example 56A

tert-Butyl - {(3S) -4-amino-3 - [(tert-butoxycarbonyl) amino] butyl} carbamate

The preparation is carried out analogously to Example 52A from 171 mg (0.52 mmol) tert-butyl - {(3S) -4-azido-3 - [(tert-butoxycarbonyl) amino] butyl} carbamate (Example 55A) in 10 ml of ethanol with the addition of 20 mg pal Ladium on activated charcoal (10%). The product is reacted without further purification.
Yield: 117 mg (75% of theory)
MS (ESI): m / z = 304 (M + H) ⁺ ; 326 (M + Na) ⁺ .

Example 57A

Benzyl - {(1S) -4 - [(tert-butoxycarbonyl) amino] -1 - [({2 - [(tert-butoxycarbonyl) amino] ethyl} amino) carbonyl] butyl} carbamate

Under argon, 300 mg (0.82 mmol) of N ² - [(benzyloxy) carbonyl] -N ⁵ - (tert-butoxycarbonyl) -L-ornithine and 171 mg (1.06 mmol) of tert-butyl (2-aminoethyl) carbamate in 6 ml of dimethylformamide dissolved. At 0 ° C (ice bath) then 204 mg (1.06 mmol) EDC and 33 mg (0.25 mmol) HOBt are added. It is slowly warmed to RT and stirred for 12 h at RT. The solution is concentrated in vacuo and the residue is taken up in ethyl acetate. The organic phase is washed successively with saturated sodium bicarbonate and sodium chloride solution, dried over magnesium sulfate and evaporated in vacuo. The remaining solid is dried under high vacuum.
Yield: 392 mg (94% of theory)
LC-MS (Method 5): R _t = 2.36 min.
MS (EI): m / z = 509 (M + H) ⁺

Example 58A

Benzyl - {(1S) -4 - [(tert-butoxycarbonyl) amino] -1 - [({3 - [(tert-butoxycarbonyl) amino] -2-hydroxypropyl} amino) carbonyl] butyl} carbamate

The preparation is analogous to Example 57A from 300 mg (0.82 mmol) of N ² - [(benzyloxy) carbonyl] -N ⁵ - (tert-butoxycarbonyl) -L-ornithine and 202 mg (1.06 mmol) of tert-butyl (3 amino-2-hydroxypropyl) carbamate in 6 ml of dimethylformamide with the addition of 204 mg (1.06 mmol) of EDC and 33 mg (0.25 mmol) of HOBt.
Yield: 412 mg (93% of theory)
LC-MS (Method 6): R _t = 2.23 min.
MS (EI): m / z = 539 (M + H) ⁺

Example 59A

N ⁵ - (tert-butoxycarbonyl) -N- {2 - [(tert-butoxycarbonyl) amino] ethyl} -L-ornithinamide

The preparation is carried out analogously to Example 52A from 390 mg (0.77 mmol) of benzyl - {(1S) -4 - [(tert-butoxycarbonyl) amino] -1 - [({2 - [(tert-butoxycarbonyl) amino] ethyl} amino) Carbonyl] butyl} carbamate (Example 57A) in 50 ml of ethanol with the addition of 40 mg of palladium on activated carbon (10%). The product is reacted without further purification.
Yield: 263 mg (91% of theory)
MS (ESI): m / z = 375 (M + H) ⁺ ; 397 (M + Na) ⁺ .

Example 60A

N ⁵ - (tert-butoxycarbonyl) -N- {3 - [(tert-butoxycarbonyl) amino] -2-hydroxypropyl} -L-ornithinamide

The preparation is carried out analogously to Example 52A from 412 mg (0.76 mmol) of benzyl - {(1S) -4 - [(tert-butoxycarbonyl) amino] -1 - [({3 - [(tert-butoxycarbonyl) amino] -2-hydroxypropyl } amino) -carbonyl] -butyl} carbamate (Example 58A) in 50 ml of ethanol with the addition of 41 mg of palladium on activated carbon (10%). The product is reacted without further purification.
Yield: 306 mg (99% of theory)
MS (ESI): m / z = 405 (M + H) ⁺ .

Example 61A

tert-Butyl - [(7R, 8S, 11S, 14S) -8 - [({(2S) -2,5-bis [(tert-butoxycarbonyl) amino] pentyl} amino) carbonyl] -11- {3- [(tert-butoxycarbonyl) amino] propyl} -5,7,17-trihydroxy-10,13-dioxo-9,12-diazatricyclo [14.3.1.1 ^2,6 ] henicosa-1 (20), 2 (21), 3,5,16,18-hexaene-14-yl] carbamate

Under argon, 23 mg (0.034 mmol) of (7R, 8S, 11S, 14S) -14 - [(tert-butoxycarbonyl) amino] -11- {3 - [(tert-butoxycarbonyl) amino] propyl} -5.7, 17-trihydroxy-10,13-dioxo-9,12-diazatricyclo [14.3.1.1 ^2,6 ] henicosa-1 (20), 2 (21), 3,5,16,18-hexaene-8-carboxylic acid ( Example 48A) and 11.9 mg (0.07 mmol) of tert -butyl {(4S) -5-amino-4 - [(tert-butoxycarbonyl) amino] pentyl} carbamate (Example 52A) dissolved in 0.75 ml of dimethylformamide. At 0 ° C (ice bath) then 8.5 mg (0.051 mmol) EDC and 1.4 mg (0.01 mmol) HOBt are added. It is slowly warmed to RT and stirred for 12 h at RT. The solution is concentrated in vacuo and the residue is stirred with water. The remaining solid is filtered off with suction and purified by preparative RP-HPLC (Reprosil ODS-A, eluent acetonitrile / 0.2% aqueous trifluoroacetic acid 5:95 → 95: 5).
Yield: 15 mg (45% of theory)
LC-MS (Method 6): R _t = 2.48 min.
MS (EI): m / z = 972 (M + H) ⁺

Analogous The procedure of Example 61A is set forth in the following table listed Examples 62A to 66A made.

Example 67A

(3S) -3 - {[(benzyloxy) carbonyl] amino} -6 - [(tert-butoxycarbonyl) amino] hexanoylmethylcarbonat

Under Argon is 2 g (5.26 mmol) of (3S) -3 - {[(benzyloxy) carbonyl] amino} -6 - [(tert-butoxycarbonyl) amino] hexanoic acid and Dissolved 0.56 g (5.73 mmol) of triethylamine in 30 ml of THF and cooled to 0 ° C. There are 0.59 g (5.73 mmol) of methyl chloroformate and left stirring for 3 hours at 0 ° C. The Reaction mixture is over Diatomaceous earth filtered. The filtrate is reacted directly.

Example 68A

Benzyl - [(1S) -4 - [(tert-butoxycarbonyl) amino] -1- (2-hydroxyethyl) butyl] carbamate

The filtrate of (3S) -3 - {[(benzyloxy) carbonyl] amino} -6 - [(tert-butoxycarbonyl) amino] hexanoylmethyl carbonate (Example 67A) becomes a suspension of 0.49 g (13.14 mmol) of sodium borohydride in 0.6 ml of water added dropwise at 0 ° C. The mixture warms slowly to room temperature and is stirred overnight. The reaction solution is concentrated in vacuo and the residue is treated with ethyl acetate and water to work up. The organic phase is dried over magnesium sulfate, concentrated in vacuo and dried under high vacuum. The crude product is reacted without further purification.
Yield: 570 mg (30% of theory)
LC-MS (Method 6): R _t = 2.09 min.
MS (EI): m / z = 367 (M + H) ⁺ .

Example 69A

tert-Butyl - [(4S) -4-amino-6-hydroxyhexyl] carbamate

The preparation is carried out analogously to Example 52A from 620 mg (1.69 mmol) of benzyl - [(1S) -4 - [(tert-butoxycarbonyl) amino] -1- (2-hydroxyethyl) butyl] carbamate (Example 68A) in 60 ml of ethanol Addition of 100 mg of palladium on charcoal (10%). The product is reacted without further purification.
Yield: 370 mg (95% of theory)
¹ H-NMR (400 MHz, D ₂ O): δ = 1.2-1.6 (m, 6H), 1.4 (s, 9H), 2.6-3.0 (m, 1H), 3.0-3.2 (m, 2H), 3.7 -3.9 (m, 2H), 4.6 (br.s, 1H).

Example 70A

Benzyl {3 - [(2 - [(tert-butoxycarbonyl) amino] -1 - {[(tert-butoxycarbonyl) amino] methyl} ethyl) amino] propyl} carbamate

310 mg (1.07 mmol) of di-tert-butyl (2-aminopropane-1,3-diyl) biscarbamate and 222 mg (1.07 mmol) of benzyl (3-oxopropyl) carbamate are dissolved in 15 ml of dichloromethane. 334 mg (1.5 mmol) sodium triacetoxyborohydride are added and stirred at room temperature overnight. The mixture is evaporated and the residue purified by preparative HPLC.
Yield: 168 mg (38% of theory)
LC-MS (Method 4): R _t = 1.76 min.
MS (EI): m / z = 481 (M + H) ⁺ .

Example 71A

Benzyl {3 - [(tert-butoxycarbonyl) (2 - [(tert-butoxycarbonyl) amino] -1 - {[(tert-butoxycarbonyl) amino] methyl} ethyl) amino] propyl} carbamate

To a solution of 168 mg (0.35 mmol) of benzyl {3 - [(2 - [(tert-butoxycarbonyl) amino] -1 - {[(tert-butoxycarbonyl) amino] methyl} ethyl) amino] propyl} carbamate (Example 70A) in 2 ml of acetonitrile is added 0.55 ml of a 10% triethylamine solution in acetonitrile and 154 mg (0.70 mmol) of di-tert-butyl dicarbonate. The reaction mixture is stirred at 60 ° C for 6 hours. The solution is concentrated and the crude product is reacted without further purification.
Yield: quant.
LC-MS (method 5): R _t = 2.87 min.
MS (EI): m / z = 580 (M + H) ⁺ .

Example 72A

Di-tert-butyl {2 - [(3-aminopropyl) (tert-butoxycarbonyl) amino] propane-1,3-diyl} biscarbamate

A solution of 190 mg (0.327 mmol) of benzyl {3 - [(tert-butoxycarbonyl) (2 - [(tert-butoxycarbonyl) amino] -1 - {[(tert-butoxycarbonyl) amino] methyl} ethyl) amino] propyl } carbamate (Example 71A) in 50 ml of glacial acetic acid / water / ethanol 4/1/1 is hydrogenated after addition of 20 mg of palladium on activated carbon (10%) for 12 h at room temperature and atmospheric pressure. It is filtered through kieselguhr and the residue is washed with ethanol. The filtrate is concentrated to dryness in vacuo. The product is reacted without further purification.
Yield: quant.
LC-MS (Method 5): R _t = 1.71 min.
MS (EI): m / z = 447 (M + H) ⁺ .

embodiments

example 1

(7R, 8S, 11S, 14S) -14-amino-N- (2-aminoethyl) -11 - [(2R) -3-amino-2-hydroxypropyl] -5,7,17-trihydroxy-10,13- dioxo-9,12-diazatricyclo [14.3.1.1 ^2,6 ] henicosa-1 (20), 2 (21), 3,5,16,18-hexaene-8-carboxamide trihydrochloride

There are 30 mg (0.32 mmol) of benzyl - {(2R) -3 - [(7R, 8S, 11S, 14S) -8 - {[(2-aminoethyl) amino] carbonyl} -5,17-bis (benzyloxy ) -14 - {[(benzyloxy) carbonyl] amino} -7-hydroxy-10,13-dioxo-9,12-diazatricyclo [14.3.1.1 ^2,6 ] henicosa-1 (20), 2 (21), 3 , 5,16,18-hexaen-11-yl] -2-hydroxypropyl} carbamate (Example 34A) in a mixture of 10 ml of acetic acid / water / ethanol 4: 1: 1. 10 mg of palladium on activated carbon (10% strength) are added thereto, followed by hydrogenation at RT and atmospheric pressure for 15 h. The reaction mixture is filtered through prewashed diatomaceous earth, washed with ethanol and the filtrate evaporated in vacuo. The residue is mixed with 0.96 ml of 0.1N aqueous hydrochloric acid and concentrated. The residue is stirred with 10 ml of diethyl ether and decanted off. The remaining solid is dried under high vacuum.
Yield: 18 mg (97% of theory).
LC-MS (Method 7): R _t = 1.92 min
MS (EI): m / z = 531 (M-3HCl + H) ⁺ .
¹ H-NMR (400 MHz, D ₂ O): δ = 1.83-1.93 (m, 1H), 1.94-2.08 (m, 1H), 2.88 (m _c , ¹ H), 2.97-3.18 (m, 4H), 3.42-3.64 (m, 3H), 3.91 (m _c, 1H), 439 (m _c, 1H), 4.82 (m _c, 1H), 5.0 (m _c, 1H), 5:56 (m _c, 1H), 6.85 -6.97 (m, 3H), 7.27-7.46 (m, 3H).

Example 2

(7R, 8S, 11S, 14S) -14-amino-N- (2-aminoethyl) -11 - [(2R) -3-amino-2-hydroxypropyl] -5,7,17-trihydroxy-10,13- dioxo-9,12-diazatricyclo [14.3.1.1 ^2,6 ] henicosa-1 (20), 2 (21), 3,5,16,18-hexaene-8-carboxamide tri (hydrotrifluoroacetate)

The trihydrochloride (Example 1) is converted into the tri (hydrotrifluoroacetate) by preparative HPLC (Reprosil ODS-A, eluent acetonitrile / 0.2% aqueous trifluoroacetic acid 5:95 → 95: 5).
Yield: quant.
¹ H-NMR (400 MHz, D ₂ O): δ = 1.83-1.93 (m, 1H), 1.94-2.08 (m, 1H), 2.88 (m _c , ¹ H), 2.97-3.18 (m, 4H), 3.42-3.64 (m, 3H), 3.91 (m _c, 1H), 4:39 (m _c, 1H), 4.82 (m _c, 1H), 5.0 (m _c, 1H), 5:56 (m _c, 1H), 6.85 -6.97 (m, 3H), 7.27-7.46 (m, 3H).

Analogous for the prescription of Example 1 are those in the following table listed Examples 3 to 6 prepared and optionally analogously to Example 2 into the corresponding tri (hydrotrifluoroacetate).

Example 7

(7R, 8S, 11S, 14S) -14-amino-N- (2-aminoethyl) -11- (3-aminopropyl) -5,7,17-trihydroxy-10,13-dioxo-9,12-diazatricyclo [ 14.3.1.1 ^2,6 ] henicosa-1 (20), 2 (21), 3,5,16,18-hexaene-8-carboxamide tri (hydrotrifluoroacetate)

The trihydrochloride (Example 5) is converted into the tri (hydrotrifluoroacetate) by preparative HPLC (Reprosil ODS-A, eluent acetonitrile / 0.2% aqueous trifluoroacetic acid 5:95 → 95: 5).
Yield: quant.
¹ H-NMR (400 MHz, D ₂ O): δ = 1.6-1.95 (m, 4H), 2.97-3.02 (m, 3H), 3.07-3.17 (m, 2H), 3.45-3.56 (m, 3H) , 4:41 (m _c, 1H), 4.78 (m _c, 1H), 4.85 (m _c, 1H), 5.5 (m _c, 1H), 6.85-6.95 (m, 3H), 7:28 to 7:39 (m, 3H) ,

Example 8

(7R, 8S, 11S, 14S) -14-amino-11- (3-aminopropyl) -N - [(2S) -2,5-diaminopentyl] -5,7,17-trihydroxy-10,13-dioxo- 9,12-diazatricyclo [14.3.1.1 ^2,6 ] henicosa-1 (20), 2 (21), 3,5,16,18-hexaene-8-carboxamide tetrahydrochloride

In 1 ml of an ice-cooled 4N hydrogen chloride-dioxane solution are added 5.5 mg (5.7 μmol) of tert-butyl [(7R, 8S, 11S, 14S) -8 - [({(2S) -2,5-bis [( tert-butoxycarbonyl) amino] pentyl} amino) carbonyl] -11- {3 - [(tert-butoxycarbonyl) amino] propyl} -5,7,17-trihydroxy-10,13-dioxo-9,12-diazatricyclo [ 14.3.1.1 ^2,6 ] henicosa-1 (20), 2 (21), 3,5,16,18-hexaen-14-yl] carbamate (Example 61A) and stirred for one hour at RT. Then concentrated to dryness in vacuo and dried to constant weight.
Yield: 4 mg (98% of theory)
LC-MS (Method 7): R _t = 0.25 min.
MS (EI): m / z = 513 (M-4HCl + H) ⁺

Analogous to the rule of Example 8, the in the following table listed Examples 9 to 13 produced.

B. Assessment of physiological effectiveness

Used abbreviations:

AMP

Adenosine monophosphate

ATP

adenosine triphosphate

BHI medium

Brain heart infusion medium

CoA

Coenzyme A

DMSO

dimethyl sulfoxide

DTT

dithiothreitol

EDTA

ethylenediaminetetraacetic

KCl

potassium chloride

KH ₂ PO ₄

potassium

_MgSO4

magnesium sulfate

MHK

Minimal inhibitory concentration

MTP

microtiter plate

NaCl

sodium chloride

Na ₂ HPO ₄

Dinatriurnhydrogenphosphat

NH ₄ Cl

ammonium chloride

NTP

nucleotide triphosphate

PBS

Phosphate Buffered saltworks

PCR

Polymerase Chain Reaction

PEG

polyethylene glycol

PEP

phosphoenolpyruvate

Tris

Tris [hydroxymethyl) aminomethane

The In vitro activity of the compounds of the invention can be seen in the following Assays are shown:

In vitro transcription-translation with E. coli extracts

to Production of an S30 extract will be logarithmically growing Escherichia coli MRE 600 (M.Mueller; University Freiburg), washed and as described for the in In vitro transcription-translation test used (Müller, M. and Blobel, G. Proc Natl Acad Sci USA (1984) 81, pp. 7421-7425).

In addition, 1 μl cAMP (11.25 mg / ml) per 50 μl reaction mixture is added to the reaction mix of the in vitro transcription-translation test. The test batch is 105 .mu.l, wherein 5 .mu.l of the substance to be tested in 5% DMSO are submitted. The transcription template used is 1 μg / 100 μl of the plasmid pBESTLuc (Promega, Germany). After incubation for 60 min at 30 ° C., 50 μl luciferin solution (20 mM tricine, 2.67 mM MgSO ₄ , 0.1 mM EDTA, 33.3 mM DTT pH 7.8, 270 μM CoA, 470 μM luciferin, 530 μM ATP) are added and the resulting bioluminescence measured for 1 minute in a luminometer. The IC ₅₀ is the concentration of an inhibitor which leads to a 50% inhibition of the translation of firefly luciferase.

In vitro transcription-translation with S. aureus extracts

Construction of a S. aureus luciferase reporter plasmids

to Construction of a reporter plasmid which is expressed in an in vitro transcription-translation assay from S. aureus can be used, the plasmid pBESTluc (Promega Corporation, USA). The one in this plasmid in front of the firefly luciferase existing E. coli tac promoter is against the capA1 promoter with corresponding Shine-Dalgarno sequence from S. aureus exchanged. For this, primers CAPFor 5'-CGGCC-AAGCTTACTCGGATCCAGAGTTTGCAAATATACAGGGGATTATATATAATGGAAAACAAGAAAGGAAAATAGGAGGTTTATATGGAAGACGCCA-3 'and CAPRev 5'-GTCATCGTCGGGAAGACCTG-3' are used. The primer CAPFor contains the capA1 promoter, the ribosome binding site and the 5 'region of luciferase Gene. After PCR using pBESTluc as a template, a PCR product which contains the firefly luciferase gene containing the fused capA1 promoter. This will be after a Restriction with ClaI and HindIII in which also with ClaI and HindIII digested vector pBESTluc ligated. The resulting plasmid p1a can in E. coli and as a template in the S. aureus in vitro transcription-translation assay be used.

Production of S30 extracts from S. aureus

Six liters of BHI medium are inoculated with a 250 ml overnight culture of a S. aureus strain and grown at 37 ° C to an OD600nm of 2-4. The cells are harvested by centrifugation and washed in 500 ml of cold buffer A (10 mM Tris-acetate, pH 8.0, 14 mM magnesium acetate, 1 mM DTT, 1 M KCl). After renewed centrifugation, the cells are washed in 250 ml of cold buffer A with 50 mM KCl and the resulting pellets are frozen at -20 ° C for 60 min. The pellets are thawed on ice in 30 to 60 minutes and taken up to a total volume of 99 ml in buffer B (10 mM Tris-acetate, pH 8.0, 20 mM magnesium acetate, 1 mM DTT, 50 mM KCl). 1.5 ml each of lysostaphin (0.8 mg / ml) in Buffer B are introduced into 3 pre-cooled centrifuge beakers and mixed with 33 ml each of the cell suspension. The samples are incubated for 45-60 min at 37 ° C with occasional shaking before adding 150 μl of a 0.5 M DTT solution. The lysed cells are centrifuged off at 30,000 × g for 30 min at 4 ° C. The cell pellet, after uptake in Buffer B, is recentrifuged under the same conditions and the collected supernatants are pooled. The supernatants are again centrifuged under the same conditions and to the upper 2/3 of the supernatant 0.25 volume of buffer C (670 mM Tris-acetate, pH 8.0, 20 mM magnesium acetate, 7 mM Na ₃ phosphoenolpyruvate, 7 mM DTT, 5.5 mM ATP , 70 μM amino acids (complete from Promega), 75 μg pyruvate kinase (Sigma, Germany)) / ml. The samples are incubated for 30 min at 37 ° C. The supernatants are dialyzed overnight at 4 ° C against 2 l of dialysis buffer (10 mM Tris-acetate, pH 8.0, 14 mM magnesium acetate, 1 mM DTT, 60 mM potassium acetate) with a buffer change in a dialysis tube with 3500 Da exclusion. The dialysate is concentrated to a protein concentration of about 10 mg / ml by covering the dialysis tube with cold PEG 8000 powder (Sigma, Germany) at 4 ° C. The S30 extracts can be stored aliquoted at -70 ° C.

Determination of IC ₅₀ in the S. aureus in vitro transcription-translation assay

The Inhibition of protein biosynthesis of the compounds can be in one in vitro transcription-translation assay to be shown. The assay is based on cell-free transcription and translation of Firefly luciferase using the reporter plasmid p1a as a template and cell-free S30 extracts obtained from S. aureus. The activity The resulting luciferase can be detected by luminescence.

The amount of S30 extract or plasmid p1a to be used must be re-tested for each preparation in order to ensure an optimal concentration in the test. 3 μl of the substance to be tested dissolved in 5% DMSO are placed in an MTP. Subsequently, 10 μl of a suitably concentrated plasmid solution p1a are added. Subsequently, 46 .mu.l of a mixture of 23 .mu.l premix (500 mM potassium acetate, 87.5 mM Tris-acetate, pH 8.0, 67.5 mM ammonium acetate, 5 mM DTT, 50 ug folic acid / ml, 87.5 mg PEG 8000 / ml, 5 mM ATP, 1.25 mM per NTP, 20 μM per amino acid, 50 mM PEP (Na ₃ salt), 2.5 mM cAMP, 250 μg per E. coli tRNA / ml) and 23 μl of a suitable amount of S. aureus S30 extract and mixed. After incubation for 60 min at 30 ° C, 50 μl luciferin solution (20 mM Tricine, 2.67 mM MgSO ₄ , 0.1 mM EDTA, 33.3 mM DTT pH 7.8, 270 μM CoA, 470 μM luciferin, 530 μM ATP) and the resulting Bioluminescence measured for 1 min in a luminometer. The IC ₅₀ is the concentration of an inhibitor which leads to a 50% inhibition of the translation of firefly luciferase.

Determination of the minimum Inhibitory concentration (MIC)

The minimum inhibitory concentration (MIC) is the minimum concentration of antibiotic used to inhibit a test bacterium in its growth for 18-24 h. The inhibitor concentration can be determined by standard microbiological procedures using modified medium as part of an agar dilution test (see, for example, The National Committee for Clinical Laboratory Standards, Methods for dilution antimicrobial susceptibility tests for bacteria that grow aerobic, approved standard-fifth edition, NCCLS document M7-A5) [ISBN 1-56238-394-9], NCCLS, 940 West Valley Road, Suite 1400, Wayne, Pennsylvania 19087-98 USA, 2000). The bacteria are cultured on 1.5% agar plates containing 20% defibrinated horse blood. The test organisms, which are incubated overnight on Columbia blood agar plates (Becton-Dickinson), are diluted in PBS, adjusted to a bacterial count of about 5 × 10 ⁵ germs / ml and dropped on test plates (1-3 μl). The test substances contain different dilutions of the test substances (dilution stages 1: 2). The cultures are incubated at 37 ° C for 18-24 hours in the presence of 5% CO ₂ .

The lowest substance concentration at which no visible bacterial growth occurs is defined as MIC. The MIC values in μg / ml of some of the compounds according to the invention compared with a series of test microbes are listed by way of example in the table below. The connections show a graded antibacterial effect against most of the test germs.

Table A (with Comparative Example 43A (biphenomycin A))

Systemic infection with S. aureus 133

The Suitability of the compounds of the invention for the treatment of bacterial infections can be found in various Animal models are shown. In addition, the animals in general infected with a suitable virulent germ and then with the compound to be tested, which in one to the respective therapy model adapted formulation is treated. Specifically, the Suitability of the compounds of the invention for the treatment of bacterial infections in a sepsis model on mice after infection with S. aureus.

For this purpose, S. aureus 133 cells are grown overnight in BH broth (Oxoid, Germany). The overnight culture was diluted 1: 100 in fresh BH broth and turned up for 3 hours. The bacteria in the logarithmic growth phase are centrifuged off and washed twice with buffered, physiological saline solution. Thereafter, a cell suspension in saline solution with an extinction of 50 units is set on the photometer (Dr. Lange LP 2W). After a dilution step (1:15), this suspension is mixed 1: 1 with a 10% mucin suspension. 0.2 ml / 20 g mouse ip is administered from this infectious solution. This corresponds to a cell number of about 1-2 × 10 ⁶ germs / mouse. IV therapy is given 30 minutes after infection. For the infection trial female CFW1 mice are used. The survival of the animals is recorded over 6 days. The animal model is adjusted so that untreated animals die within 24 hours of infection. For example compound 2, a therapeutic effect of ED ₁₀₀ = 1.25 mg / kg could be demonstrated in this model.

Determination of spontaneous resistance frequencies against S. aureus

The spontaneous resistance rates of the compounds according to the invention are determined as follows: the bacterial germs are cultivated on blood agar plates or Müller-Hinton agar plates at 37 ° C. overnight, resuspended in 2 ml PBS (about 2 × 10 ⁹ germs / ml). 100 μl of this cell suspension or 1:10 and 1: 100 dilutions are diluted on predried agar plates (1.5% agar, 20% defibrinated horse blood or 1.5% agar, 20% bovine serum in 1/10 Müller-Hinton medium diluted with PBS), which contains the compound according to the invention to be tested in a concentration corresponding to 5 × MIC or 10 × MIC, and incubated at 37 ° C. for 48 h. The resulting colonies (cfu) are counted.

Isolation of Binhenomycin-Resistant S. aureus Strains RN4220Bi ^R and T17

The S. aureus strain RN4220Bi ^R is isolated in vitro. For this purpose, in each case 100 .mu.l of a S. aureus RN4220 cell suspension (about 1.2 × 10 ⁸ cfu / ml) on an antibiotic agar plate (18.5 mM Na ₂ HPO ₄ , 5.7 mM KH ₂ PO ₄ , 9.3 mM NH ₄ Cl, 2.8 mM MgSO ₄ , 17.1 mM NaCl, 0.033 μg / ml thiamine hydrochloride, 1.2 μg / ml nicotinic acid, 0.003 μg / ml biotin, 1% glucose, 25 μg / ml of each proteinogenic amino acid with the addition of 0.4% BH broth and 1% agarose) and an agar plate containing 2 μg / ml biphenomycin B (10x MIC) and incubated overnight at 37 ° C. While approximately 1 × 10 ⁷ cells grow on the antibiotic-free plate, approximately 100 colonies grow on the antibiotic-containing plate, corresponding to a resistance frequency of 1 × 10 ^-5 . Some of the colonies grown on the antibiotic-containing plate are tested for biphenomycin B MIC. A colony with a MIC> 50 μM is selected for further use and the strain is designated RN4220Bi ^R.

The S. aureus strain T17 is isolated in vivo. CFW1 mice are infected intraperitoneally with 4 × 10 ⁷ S. aureus 133 cells per mouse. 0.5 hours after infection, the animals are treated intravenously with 50 mg / kg biphenomycin B. The survivors are removed from the kidneys on day 3 post-infection. After homogenizing the organs, the homogenates are plated on antibiotic-free and antibiotic-containing agar plates as described for RN4220Bi ^R and incubated overnight at 37 ° C. Approximately half of the colonies isolated from the kidney show growth on the antibiotic-containing plates (2.2 × 10 ⁶ colonies), demonstrating the accumulation of biphenomycin B-resistant S. aureus cells in the kidney of the treated animals. Approximately Twenty of these colonies are tested for biphenomycin B MIC and a colony with a MIC> 50 μM is selected for further culture and the strain is designated T17.

C. Embodiments for pharmaceutical compositions

The Compounds of the invention can as follows be converted into pharmaceutical preparations:

Intravenously administrable Solution:

Composition:

1 mg of the compound of Example 1, 15 g of polyethylene glycol 400 and 250 g of water for Injections.

production:

The inventive compound is dissolved together with polyethylene glycol 400 in the water with stirring. The solution is sterile filtered (pore diameter 0.22 microns) and under aseptic conditions in heat-sterilized infusion bottles filled. These are with infusion stoppers and crimp caps locked.

Claims (11)

Compound of the formula

where R ¹ is a group of the formula

wherein R ⁷ is hydrogen or hydroxy, * is the point of attachment to the carbon atom, R ² is hydrogen, methyl or ethyl, R ³ is a group of the formula

where * is the point of attachment to the nitrogen atom, R ⁸ is hydrogen, amino or hydroxy, R ⁹ is hydrogen, methyl or a group of the formula

wherein * is the point of attachment to the nitrogen atom, R ²⁰ is hydrogen or * - (CH ₂ ) _f -NHR ²² , wherein R ²² is hydrogen or methyl and f is a number 1, 2 or 3, R ²¹ is hydrogen or methyl, d is a number 0, 1, 2 or 3 and e is a number 1, 2 or 3, R ¹⁰ is hydrogen or aminoethyl, or R ⁹ and R ¹⁰ form together with the nitrogen atom to which they are attached form a piperazine ring, R ¹¹ and R ^{1 5 are} independently * - (CH _{2) Z1} -OH, * - (CH _{2) Z2} -NHR ^16, hydroxycarbonyl, Aminocarbonyl, hydroxycarbonylmethyl, aminocarbonylmethyl, * -CONHR ¹⁷ or * -CH ₂ CONHR ^{1 8} where * is the point of attachment to the carbon atom, Z1 and Z2 are independently a number 1, 2 or 3, R ¹⁶ is hydrogen or Methyl and R ¹⁷ and R ^{18 are} independently a group of the formula

in which * is the point of attachment to the nitrogen atom, R ^8a is hydrogen, amino or hydroxy, R ^9a is hydrogen, methyl or aminoethyl, R ^10a is hydrogen or aminoethyl, or R ^9a and R ^10a form together with the nitrogen atom to which they are attached are a piperazine ring, R ^11a and R ^{1 5a are} independently * - (CH ₂ ) _{Z 1a} -OH, * - (CH ₂ ) _Z 2a -NHR ^16a , hydroxycarbonyl, aminocarbonyl, hydroxycarbonylmethyl or aminocarbonylmethyl; wherein * is the point of attachment to the carbon atom, R ^16a is hydrogen or methyl and Z1a and Z2a are independently a number 1, 2 or 3, R ^{12 a} and R ^{1 4a are} independently hydrogen or methyl, R ^13a is amino or Hydroxy is, ka is a number 0 or 1, la, wa, xa and ya are independently a number 1, 2, 3 or 4, R ¹² and R ^{14 are} independently hydrogen, methyl or a group of the formula

are wherein * is the linkage site to the nitrogen atom, R ^20a is hydrogen or * - (CH _{2) fa} -NHR ^22a, wherein R ^22a is hydrogen or methyl, and fa denotes a number 1, 2 or 3, R ^21a is hydrogen or methyl, since a number is 0, 1, 2 or 3 and ea is a number 1, 2 or 3, R ¹³ is amino or hydroxy, R ^{19 is} a group of the formula

in which * is the point of attachment to the nitrogen atom, R ^8b is hydrogen, amino or hydroxy, R ^9b is hydrogen, methyl or aminoethyl, R ^10b is hydrogen or aminoethyl, or R ^9b and R ^10b form together with the nitrogen atom to which they are attached are a piperazine ring, R ^11b and R ^{15 b are} independently * - (CH ₂ ) _{Z 1b} -OH, * - (CH ₂ ) _Z 2b -NHR ^16b , hydroxycarbonyl, aminocarbonyl, hydroxycarbonylmethyl or aminocarbonylmethyl, wherein the point of attachment to the carbon atom is, R ^16b is hydrogen or methyl and Z1b and Z2b are independently a number 1, 2 or 3, R ^{1 2b} and R ^{14b are} independently hydrogen or methyl, R ^13b is amino or hydroxy, kb is a number 0 or 1, lb, wb, xb and yb are independently a number 1, 2, 3 or 4, k and t are independently a number 0 or 1, l, w, x and y are independently one Number 1, 2, 3 or 4 are m, r, s and v are independent are mutually a number 1 or 2, n, o, p and q are independently a number 0, 1 or 2, u is a number 0, 1, 2 or 3,

or y is independently of one another l, w, x or y can be a hydroxyl group, R ⁴ is hydrogen, halogen, amino, hydroxyl, aminocarbonyl, hydroxycarbonyl, nitro, trifluoromethyl, trifluoromethoxy, (C ₁ -C ₄ ) - Alkyl, (C ₁ -C ₄ ) alkoxy or (C ₁ -C ₆ ) alkylamino, R ⁵ is hydrogen, halogen, amino, hydroxy, aminocarbonyl, hydroxycarbonyl, nitro, trifluoromethyl, trifluoromethoxy, (C ₁ -C ₄ ) -Alkyl, (C ₁ -C ₄ ) -alkoxy or (C ₁ -C ₆ ) -alkylamino, R ⁶ is hydrogen, methyl or ethyl, or one of its salts, its solvates or the solvates of its salts. Compound according to claim 1, characterized in that it has the formula

in which R ¹ to R ^{6 have} the same meaning as in formula (I), or one of their salts, their solvates or the solvates of their salts. A compound according to claim 1 or 2, characterized in that R ¹ is a group of the formula

wherein R ⁷ is hydrogen or hydroxy, * is the point of attachment to the carbon atom, R ² is hydrogen or methyl, R ³ is a group of the formula

where * is the point of attachment to the nitrogen atom, R ⁸ is hydrogen, amino or hydroxy, R ⁹ is hydrogen, methyl or a group of the formula

wherein * is the point of attachment to the nitrogen atom, R ²⁰ is hydrogen or * - (CH ₂ ) _f -NHR ²² , wherein R ²² is hydrogen and f is a number 1, 2 or 3, R ²¹ is hydrogen d is a number 0 and e is a number 1, 2 or 3, R ¹⁰ is hydrogen or aminoethyl, R ¹¹ * is - (CH _{2) Z1} -OH, * - (CH _{2) Z2} -NHR ^{1 6,} hydroxycarbonyl, aminocarbonyl, hydroxycarbonylmethyl, aminocarbonyl-methyl, * -CONHR ¹⁷ or * -CH ₂ CONHR ^18, wherein * the linkage site to the carbon atom, Z1 and Z2 independently of one another are a number 1, 2 or 3, R ^{1 6} is hydrogen or methyl and R ¹⁷ and R ¹⁸ independently of one another a group of formula

in which * is the point of attachment to the nitrogen atom, R ^8a is hydrogen, amino or hydroxy, R ^9a is hydrogen, methyl or aminoethyl, R ^10a is hydrogen or aminoethyl, or R ^9a and R ^10a form together with the nitrogen atom to which they are attached are a piperazine ring, R ^11a and R ^{1 5a are} independently * - (CH ₂ ) _{Z 1a} -OH, * - (CH ₂ ) _Z 2a -NHR ^{1 6a} , hydroxycarbonyl, aminocarbonyl, hydroxycarbonylmethyl or aminocarbonylmethyl in which * is the point of attachment to the carbon atom, R ^{1 6a} is hydrogen or methyl and Z1a and Z2a are independently a number 1, 2 or 3, R ^{1 2a} and R ^{1 4a are} independently hydrogen or methyl, R ^{13a is the} same Amino or hydroxy, ka is a number 0 or 1, la, wa, xa and ya independently of one another are a number 1, 2, 3 or 4, R ¹² is hydrogen or methyl, k is a number 0 or 1, l and w are independently of one another a number 1, 2, 3 or 4,

or w independently of each other at l or w equal to 3 can carry a hydroxy group, R ⁴ is hydroxy, R ⁵ is hydrogen, R ⁶ is hydrogen. Process for the preparation of a compound of formula (I) according to claim 1 or one of its salts, solvates or solvates of its salts, characterized in that [A] is a compound of formula

wherein R ¹ , R ² , R ⁴ , R ⁵ and R ^{6 have} the meaning given in claim 1 and boc is tert-butoxycarbonyl, in a two-stage process, first in the presence of one or more dehydrating reagents with a compound of formula H ₂ NR ³ (III), wherein R ^{3 has} the meaning given in claim 1, and then reacted with an acid, or [B] a compound of the formula

wherein R ¹ , R ² , R ⁴ , R ⁵ and R ^{6 have} the meaning given in claim 1 and Z is benzyloxycarbonyl, in a two-stage process, first in the presence of one or more dehydrating reagents with a compound of formula H ₂ NR ³ (III), wherein R ^{3 has} the meaning given in claim 1, and then reacted with an acid or by hydrogenolysis, or [C] a compound of the formula

wherein R ¹ , R ² , R ⁴ , R ⁵ and R ^{6 have} the meaning given in claim 1 and Z is benzyloxycarbonyl and R ²³ is benzyl, methyl or ethyl, in a two-step process, first in the presence of potassium cyanide with a compound of the formula H ₂ NR ³ (III), wherein R ^{3 has} the meaning given in claim 1, and then reacted with an acid or by hydrogenolysis. Process for the preparation of a compound of the formula (I) according to claim 1 or one of its solvates, characterized that is a salt of the compound or a solvate of a salt of the compound by chromatography with the addition of a base in the compound or one of their solvates is transferred. A compound according to any one of claims 1 to 3 for the treatment and / or prophylaxis of diseases. Use of a compound according to any one of claims 1 to 3 for the manufacture of a medicament for the treatment and / or prophylaxis of diseases. Use of a compound according to any one of claims 1 to 3 for the manufacture of a medicament for the treatment and / or prophylaxis of bacterial diseases. Medicaments containing at least one compound according to one of the claims 1 to 3 in combination with at least one inert, non-toxic, pharmaceutically suitable excipient. Medicament according to claim 9 for the treatment and / or Prophylaxis of bacterial infections. Method of combating bacterial infections in humans and animals by administering an antibacterial effective amount of at least one compound according to any one of claims 1 to 3 or a medicament according to claim 9.