Vancomycin derivative, and preparation method and application thereof
09828413 · 2017-11-28
Assignee
Inventors
Cpc classification
A61K38/14
HUMAN NECESSITIES
C07K9/008
CHEMISTRY; METALLURGY
International classification
A61K38/14
HUMAN NECESSITIES
Abstract
The present invention provides a vancomycin derivative, and a preparation method and an application thereof. The vancomycin derivative of the present invention is obtained by introducing a glycerate moiety between a vancomycin derivative and a liposoluble modification group and has reduced liposolubility and improved water solubility, thereby reducing a side effect in the cardiovascular aspect.
Claims
1. A vancomycin derivative of formula (I): ##STR00084## wherein: R.sup.1 is —NHCH.sub.3 or —NH.sub.2; R.sup.2 is H or 4-epi-vancosaminyl; R.sup.3 is —(R)COOR.sup.a or —(S)COOR.sup.a or —(R/S)COOR.sup.a; wherein R.sup.a is H, C1-C20 alkyl, C5-C12 aryl, C2-C12 alkenyl or C2-C12 alkynyl; R.sup.4 is hydrogen, C1-C20 alkyl, C5-C12 aryl, C2-C12 alkenyl, C2-C12 alkynyl, (C1-C20 alkyl)- R.sup.5 or (C1-C20 alkyl)-O—R.sup.5; wherein R.sup.5 has the following structure: (a) unsubstituted C5-C12 aryl or mono-substituted C5-C12 aryl or poly-substituted C5-C12 aryl, wherein the substituent independently is: (I) hydroxyl (II) halogen (III) nitro (IV) amino (V) C1-C20 alkyl (b) the following structure: ##STR00085## A.sup.1 is —OC(A.sup.2)2-C(A.sup.2)2-O— or —O—C(A.sup.2)2-O— or —C(A.sup.2)2-O— or —C(A.sup.2)2-C(A.sup.2)2-C(A.sup.2)2-C(A.sup.2)2-, wherein A.sup.2 independently is hydrogen or C1-C20 alkyl (c) the following structure: ##STR00086## q is 0-4, wherein R.sup.7 independently is the following group: (I) hydrogen (II) hydroxyl (III) halogen (IV) nitro (V) amino (VI) C1-C20 alkyl r is 1-5, but q+r is no more than 5 Z is the following case: (I) a single bond (II) —(C1-C12)alkyl- R.sup.8 independently is: (I) C5-C12 aryl (II) C5-C12 heteroaryl (III) phenyl unsubstituted or substituted with 1 to 5 substituents independently selected from: (a) hydrogen (b) hydroxyl (c) halogen (d) nitro (e) amino (f) C1-C20 alkyl.
2. The vancomycin derivative of claim 1, wherein: R.sup.1 is —NHCH.sub.3 or —NH.sub.2; R.sup.2 is H or 4-epi-vancosaminyl; R.sup.3 is —(R)COOR.sup.a or —(S)COOR.sup.a or —(R/S)COOR.sup.a; wherein R.sup.a is H, C1-C20 alkyl, C5-C12 aryl, C2-C12 alkenyl or C2-C12 alkynyl; R.sup.4 is C1-C20 alkyl.
3. The vancomycin derivative of claim 1, wherein: R.sup.1 is —NHCH.sub.3 or —NH.sub.2; R.sup.2 is H or 4-epi-vancosaminyl; R.sup.3 is —(R)COOR.sup.a or —(S)COOR.sup.a or —(R/S)COOR.sup.a; wherein R.sup.a is H, C1-C20 alkyl, C5-C12 aryl, C2-C12 alkenyl or C2-C12 alkynyl; R.sup.4 is (C1-C20 alkyl)-R.sup.5, wherein R.sup.5 has the following structure: ##STR00087## p is 1-5, wherein R.sup.7 independently is the following group: (I) hydrogen (II) hydroxyl (III) halogen (IV) nitro (V) amino (VI) C1-C20 alkyl.
4. The vancomycin derivative of claim 1, wherein: R.sup.1 is —NHCH.sub.3 or —NH.sub.2; R.sup.2 is H or 4-epi-vancosaminyl; R.sup.3 is —(R)COOR.sup.a or —(S)COOR.sup.a or —(R/S)COOR.sup.a; wherein R.sup.a is H, C1-C20 alkyl, C5-C12 aryl, C2-C12 alkenyl or C2-C12 alkynyl; R.sup.4 is (C1-C20 alkyl)-R.sup.5, wherein R.sup.5 has the following structure: ##STR00088## q is 0-4, wherein R.sup.7 independently is the following group: (I) hydrogen (II) hydroxyl (III) halogen (IV) nitro (V) amino (VI) C1-C20 alkyl r is 1-5, but q+r is no more than 5 Z is the following case: (I) a single bond (II) —(C1-C12)alkyl- R.sup.8 independently is: (I) C5-C12 aryl (II) C5-C12 heteroaryl (III) phenyl unsubstituted or substituted with 1 to 5 substituents independently selected from: (a) hydrogen (b) hydroxyl (c) halogen (d) nitro (e) amino (f) C1-C20 alkyl.
5. The vancomycin derivative of claim 1, wherein: R.sup.1 is —NHCH.sub.3 or —NH.sub.2; R.sup.2 is H or 4-epi-vancosaminyl; R.sup.3 is —(R)COOR.sup.a or —(S)COOR.sup.a or —(R/S)COOR.sup.a; wherein R.sup.a is H; R.sup.4 is (C1-C20 alkyl)-R.sup.5, wherein R.sup.5 has the following structure: ##STR00089## q is 0-4, wherein R.sup.7 independently is the following group: (I) hydrogen (II) hydroxyl (III) halogen (IV) nitro (V) amino (VI) C1-C20 alkyl r is 1-5, but q+r is no more than 5 Z is the following case: (I) a single bond (II) —(C1-C12)alkyl- R.sup.8 independently is: (I) C5-C12 aryl (II) C5-C12 heteroaryl (III) phenyl unsubstituted or substituted with 1 to 5 substituents independently selected from: (a) hydrogen (b) hydroxyl (c) halogen (d) nitro (e) amino (f) C1-C20 alkyl.
6. A method for treating an infection caused by a gram-positive bacteria or vancomycin-resistant bacteria comprising contacting the bacteria with a compound as described in claim 1.
7. A process for preparing the vancomycin derivative according to claim 1, comprising treating vancomycin or an analogue thereof and a compound of formula ##STR00090## with a reductive agent in a polar solvent followed by hydrolysis, and if R.sup.a is H in the formula, the product is directly obtained after reduction; wherein the vancomycin and the analogue thereof are vancomycin of formula (II), norvancomycin of formula (III), 4-epi-vancosaminyl vancomycin of formula (IV) or 4-epi-vancosaminyl norvancomycin of formula (V): ##STR00091## wherein M is alkali metal or alkaline earth metal; R.sup.3 is —(R)COOR.sup.a or —(S)COOR.sup.a or —(R/S)COOR.sup.a, and R.sup.a is H, C1-C20 alkyl, C5-C12 aryl, C2-C12 alkenyl or C2-C12 alkynyl; R.sup.4 is hydrogen, C1-C20 alkyl, C5-C12 aryl, C2-C12 alkenyl, C2-C12 alkynyl, (C1-C20 alkyl)- R.sup.5 or (C1-C20 alkyl)-O—R.sup.5, and R.sup.5 has the following structure: (a) unsubstituted C5-C12 aryl or mono-substituted C5-C12 aryl or poly-substituted C5-C12 aryl, wherein the substituent independently is: (I) hydroxyl (II) halogen (III) nitro (IV) amino (V) C1-C20 alkyl (b) the following structure: ##STR00092## A.sup.1 is —OC(A.sup.2)2-C(A.sup.2)2-O— or —O—C(A.sup.2)2-O— or —C(A.sup.2)2-N— or —C(A.sup.2)2-C(A.sup.2)2-C(A.sup.2)2-C(A.sup.2)2-, wherein A.sup.2 independently is hydrogen or C1-C20 alkyl (c) the following structure: ##STR00093## q is 0-4, wherein R.sup.7 independently is the following group: (I) hydrogen (II) hydroxyl (III) halogen (IV) nitro (V) amino (VI) C1-C20 alkyl r is 1-5, but q+r is no more than 5 Z is the following case: (I) a single bond (II) —(C1-C12)alkyl- R.sup.8 independently is: (I) C5-C12 aryl (II) C5-C12 heteroaryl (III) phenyl unsubstituted or substituted with 1 to 5 substituents independently selected from: (a) hydrogen (b) hydroxyl (c) halogen (d) nitro (e) amino (f) C1-C20 alkyl.
8. The process according to claim 7, wherein the polar solvent is methanol, ethanol, iso-propanol, tert-butanol, N, N-dimethylformamide, or N, N-dimethylacetamide; the temperature is between 0 and 80° C.; and the reductive agent is sodium borohydride, potassium borohydride, borane or a complex containing borane, sodium cyano borohydride, potassium cyano borohydride, sodium triacetoxy borohydride, or potassium triacetoxy borohydride.
9. A method for treating an infection caused by a gram-positive bacteria or vancomycin-resistant bacteria in an animal comprising, administering a compound as described in claim 1 to the animal.
10. The vancomycin derivative of claim1 that is selected from the group consisting of: ##STR00094## ##STR00095## ##STR00096## ##STR00097## ##STR00098## ##STR00099## ##STR00100## ##STR00101## ##STR00102## ##STR00103## ##STR00104## ##STR00105## ##STR00106## ##STR00107## ##STR00108## ##STR00109## ##STR00110## ##STR00111##
11. The vancomycin derivative of claim 1, wherein: R.sup.1 is —NHCH.sub.3 or —NH.sub.2; R.sup.2 is H or 4-epi-vancosaminyl; R.sup.3 is —(R)COOR.sup.a or —(S)COOR.sup.a or —(R/S)COOR.sup.a; wherein R.sup.a is H, C1-C20 alkyl, C5-C12 aryl, C2-C12 alkenyl or C2-C12 alkynyl; R.sup.4 is hydrogen, C1-C20 alkyl, C5-C12 aryl, C2-C12 alkenyl, C2-C12 alkynyl, (C1-C20 alkyl)-R.sup.5 or (C1-C20 alkyl)-O-R.sup.5; and R.sup.5 has the following structure: ##STR00112## wherein, p is 1-5; and each R.sup.7 is independently: (I) hydroxyl (II) halogen (III) nitro (IV) amino or (V) C1-C20 alkyl.
12. The process of claim 7, wherein R.sup.5 has the following structure: ##STR00113## wherein p is 1-5; and each R.sup.7 is independently: (I) hydroxyl (II) halogen (III) nitro (IV) amino or (V) C1-C20 alkyl.
Description
DETAILED DESCRIPTION OF THE INVENTION
(1) In Vitro Activity Assay
(2) The compound of formula 1 of the present invention or a clinically acceptable salt thereof is intended to be used for treatment of gram-positive bacteria or vancomycin-resistant bacteria infection cases.
(3) To verify the activity, a group of the compounds of the present invention were preferably subjected to in vitro activity assay (Table 1).
(4) TABLE-US-00001 TABLE 1 The compounds of formula (I) No. Structure V9
(5) In vitro activity assay was performed according to Microbiological Identification of Antibiotics, Appendix XIA, Volume II, Chinese Pharmacopoeia 2010. Vancomycin-sensitive Staphylococcus aureus strains (Newman and Mu 50) were selected as the test strains, and trypticase soy broth was selected as the culture medium. The assay for minimum inhibitory concentration (MIC) was performed as follows: the compound to be tested was dissolved in N,N-dimethylformamide to prepare a stock solution at 1.28 mg/ml, the stock solution was diluted with the culture medium to a initial concentration of 1.28 μg/ml, which was subsequently half diluted to prepare test solutions at 64 μg/ml-0.125 μg/ml, and the assay was performed according to Cup-Plate Method, Microbiological Identification of Antibiotics, Appendix XIA, Volume II, Chinese Pharmacopoeia 2010, wherein vancomycin and blank were used as controls. The results of in vitro activity assay of the compounds of formula (I) are listed in Table 2.
(6) TABLE-US-00002 TABLE 2 MIC values (μg/ml) Test strains Staphylococcus Compounds Staphylococcus aureus Newman aureus Mu50 V9 8 32 V11 8 32 V13 <0.125 2 V15 <0.125 2 V16 16 64 V19 64 >128 V20 <0.125 2 V21 <0.125 2 V22 2 8 V23 2 8 V24 4 8 V25 4 8 V26 16 64 V27 16 64 V30 4 16 V31 2 8 V32 2 8 V33 16 64 V51 8 32 V52 <0.125 2 V53 <0.125 2 V54 <0.125 2 V55 <0.125 2 V57 4 16 V58 64 >128 V59 2 8 V60 2 8 V61 4 8 V62 4 8 V63 <0.125 2 V64 <0.125 2 V65 <0.125 2 V66 <0.125 2 V67 2 4 V68 4 8 V69 2 8 DMSO >128 >128 Vancomycin 2 8
(7) It is seen from the results that each group of the compounds exhibited varying degrees of antibacterial activity against vancomycin-sensitive Staphylococcus aureus strains. With increase in liposolubility of the group R.sup.5, there is a trend in which the inhibitory activity of the compounds against the bacteria is enhanced.
(8) Solubility Test of Compounds
(9) Solubility test of each compound was performed according to the guidelines of General Notices, Volume II, Chinese Pharmacopoeia 2005: weigh out finely powdered compound, place the compound in different volumes of water, strongly shake for 30 seconds at an interval of 5 minutes; observe the solubility behavior within 30 minutes, and obtain the solubility range of the compound, wherein all the solubility data range are measured at a temperature of 25° C. Solubility of vancomycin and the analogues thereof are listed in Table 3.
(10) TABLE-US-00003 TABLE 3 Solubility of the compounds in water Solubility in water Compounds (mg/ml) Vancomycin ≧100 Oritavancin <0.1 (data from US2010/045201) V9 <0.1 V11 50-60 V13 50-60 V15 ≧60 V16 ≧60 V19 50-60 V20 <5 V21 >8 V22 <5 V23 >8 V24 >10 V25 <5 V26 4.5 V27 20 V30 4 V31 <1 V32 50-60 V33 5 V51 20 V52 20 V53 20 V54 15-20 V55 3 V57 >60 V58 50-60 V59 <10 V60 >20 V61 20 V62 5-20 V63 5-20 V64 5-20 V65 5-20 V66 5-20 V67 5-10 V68 5-10 V69 5-10
(11) It is seen from the solubility data that after introducing a glycerate moiety into the structure, the solubility of the compound in water increases by 1-2 orders of magnitude as compared to Oritavancin. This result demonstrates that the glycerate moiety plays a critical role in increasing the solubility in water.
(12) Preparation Process
(13) Provided is a preparation process, which is a process for preparing the vancomycin derivative according to any one of claims 1-5:
(14) ##STR00063## and in which the product is obtained from reductive reaction of vancomycin or an analogue thereof and a compound of formula
(15) ##STR00064## with a reductive agent in a polar solvent followed by hydrolysis, and if R.sup.a is H in the formula, the product is directly obtained after reduction without further hydrolysis; specifically, the reaction is performed as follows:
(16) ##STR00065##
(17) The present invention is further illustrated by the following examples, which should not be construed as limiting the present invention.
Example 1
(18) ##STR00066## ##STR00067##
Synthetic Procedure:
Step 1:
(19) ##STR00068##
(20) A 500 ml single necked flask was charged with 2.19 g of sodium hydride, suspended with 100 ml of N,N-dimethylformamide, cooled to 0-5° C. under nitrogen atmosphere, 10.0 g of 4-chlorophenyl benzyl alcohol was dissolved in 100 ml of N,N-dimethylformamide and was added to the reaction solution dropwise slowly, and after addition, the reaction was stirred for 0.5 hour followed by addition of 7.6 g of ethyl bromoacetate, and after addition, the temperature was raised to 35-40° C. overnight, and after the reaction completed as shown by TLC, the reaction was poured into 1 L of ice-water and was added with 500 ml of ethyl acetate for extraction, the organic phase was washed with saturated sodium chloride, dried over anhydrous sodium sulfate and then concentrated to dryness by a rotary evaporator to obtain a crude product, which was purified by column eluted with 10% ethyl acetate/petroleum ether to obtain 11.0 g of an oily liquid with a yield of 83.0%.
(21) Step 2:
(22) ##STR00069##
(23) A 100 ml single necked flask was charged with 2.5 g of potassium tert-butoxide, dispersed with 15 ml of diethyl ether, a solution of 5.9 g of the product obtained from the previous step in 2.2 ml of methyl formate was added slowly under nitrogen atmosphere, the reaction solution was reacted at room temperature overnight, and after the reaction completed as shown by TLC, 50 ml of diethyl ether was added and stirred for 0.5 hour followed by suction filtration, the filter cake was dried under reduced pressure to obtain 5.6 g of a white solid.
(24) Step 3:
(25) ##STR00070##
(26) A 100 ml single necked flask was charged with 743 mg of vancomycin, which was dissolved in 40 ml of N,N-dimethylformamide at 80° C., 214 mg of the product obtained from the previous step was added, followed by addition of 63 mg of sodium cyano borohydride in batch, and after addition, the reaction was performed for 2 hours, 1 ml of acetic acid was added and stirred for 0.5 hour, the reaction solution was poured into 50 ml of diethyl ether whereupon a solid precipitated, suction filtration was performed, the filter cake was stirred/washed with 40 ml of a solvent mixture of methanol and diethyl ether (1:3) followed by suction filtration, the crude product thus obtained was isolated by preparative HPLC to obtain 100 mg of the product. MS m/e 1750.4, 1751.4, 1752.4 (M+1)
(27) Step 4:
(28) ##STR00071##
(29) 30 mg of the product obtained from the previous step was dissolved in a solvent mixture of 3 ml of tetrahydrofurane and 3 ml of water, 4.6 mg of lithium hydroxide was added with stirring, the reaction solution was stirred for 4 hours, 18 mg of acetic acid was added to quench the reaction, the organic solvent was removed by a rotary evaporator, purification by preparative HPLC obtained 9.7 mg of the product, MS m/e 1736.5, 1738.5, 1739.5 (M+1)
Example 2
(30) Compounds V9, V11, V13, V15, V20, V21, V22, V23, V24, V25, V55, V61 and the like were prepared according to the process as described in Example 1.
Example 3
(31) ##STR00072## ##STR00073##
Synthetic Procedure:
Step 1:
(32) ##STR00074##
(33) A 100 ml single necked flask was charged with 20 ml of n-butanol, 1.80 g of pieces of sodium was added in an ice-water bath, and after addition, the mixture was heated at reflux until the solid dissolved, cooled to room temperature, 10.0 g of ethyl bromoacetate was added, after which the temperature was raised to 40-50° C., stirred overnight, and after the reaction completed as shown by TLC, 100 ml of diethyl ether was added, the mixture was washed with 50 ml of water three times, the organic phase was dried by a rotary evaporator under reduced pressure to obtain 9.1 g of an oily liquid, which was directly used in the next step.
(34) Step 2:
(35) ##STR00075##
(36) A 100 ml single necked flask was charged with 2.5 g of potassium tert-butoxide, dispersed with 15 ml of diethyl ether, a solution of 3.0 g of the product obtained from the previous step in 2.2 ml of methyl formate was added slowly under nitrogen atmosphere, the reaction solution was reacted at room temperature overnight, and after the reaction completed as shown by TLC, 50 ml of diethyl ether was added and stirred for 0.5 hour followed by suction filtration, the filter cake was dried under reduced pressure to obtain 2.9 g of a white solid.
(37) Step 3:
(38) ##STR00076##
(39) A 250 ml single necked flask was charged with 1.48 g of vancomycin, which was dissolved in 80 ml of N,N-dimethylformamide at 80° C., 276 mg of the product obtained from the previous step was added, followed by addition of 126 mg of sodium cyano borohydride in batch, and after addition, the reaction was performed for 2 hours, 5 ml of acetic acid was added and stirred for 0.5 hour, the reaction solution was poured into 100 ml of diethyl ether whereupon a solid precipitated, suction filtration was performed, the filter cake was stirred/washed with 40 ml of a solvent mixture of methanol and diethyl ether (1:3) followed by suction filtration, the crude product thus obtained was isolated by preparative HPLC to obtain 56 mg of the product. MS m/e 1606.5, 1607.5, 1608.5 (M+1)
(40) Step 4:
(41) ##STR00077##
(42) 30 mg of the product obtained from the previous step was dissolved in a solvent mixture of 3 ml of tetrahydrofurane and 3 ml of water, 7.8 mg of lithium hydroxide was added with stirring, the reaction solution was stirred for 4 hours, 18 mg of acetic acid was added to quench the reaction, the organic solvent was removed by a rotary evaporator, purification by preparative HPLC obtained 5.0 mg of the product, MS m/e 1592.2, 1593.2 (M+1)
Example 4
(43) Compounds V16, V19, V26, V27, V30, V31, V32, V33, V67, V68 and the like were prepared according to the process as described in Example 1.
Example 5
(44) ##STR00078##
Synthetic Procedure:
Step 1:
(45) ##STR00079##
(46) A 250 ml single necked flask was charged with 1.5 g of norvancomycin, which was dissolved in 80 ml of N,N-dimethylformamide at 80° C., 250 mg of the product obtained from Step 2 of Example 1 was added, followed by addition of 130 mg of sodium cyano borohydride in batch, and after addition, the reaction was performed for 2 hours, 5 ml of acetic acid was added and stirred for 0.5 hour, the reaction solution was poured into 100 ml of diethyl ether whereupon a solid precipitated, suction filtration was performed, the filter cake was stirred/washed with 40 ml of a solvent mixture of methanol and diethyl ether (1:3) followed by suction filtration, the crude product thus obtained was isolated by preparative HPLC to obtain 15 mg of the product. MS m/e 1736.5, 1737.5, 1738.5 (M+1)
(47) Step 2:
(48) ##STR00080##
(49) 5 mg of the product obtained from the previous step was dissolved in a solvent mixture of 1 ml of tetrahydrofuran and 1 ml of water, 2.0 mg of lithium hydroxide was added with stirring, the reaction solution was stirred for 1 hour, 10 mg of acetic acid was added to quench the reaction, the organic solvent was removed by a rotary evaporator, purification by preparative HPLC obtained 3.5 mg of the product, MS m/e 1722.5, 1723.5, 1724.5 (M+1)
Example 6
(50) Compounds V51, V52, V53, V54, V55, V57, V58, V59, V60, V68 and the like were prepared according to the process as described in Example 1.
Example 7
(51) ##STR00081##
Synthetic Procedure:
Step 1:
(52) ##STR00082##
(53) A 500 ml single necked flask was charged with 3.1 g of 4-epi-vancosaminyl vancomycin, which was dissolved in 150 ml of N,N-dimethylformamide at 80° C., 500 mg of the product obtained from Step 2 of Example 1 was added, followed by addition of 250 mg of sodium cyano borohydride in batch, and after addition, the reaction was performed for 2 hours, 7 ml of acetic acid was added and stirred for 0.5 hour, the reaction solution was poured into 150 ml of diethyl ether whereupon a solid precipitated, suction filtration was performed, the filter cake was stirred/washed with 40 ml of a solvent mixture of methanol and diethyl ether (1:3) followed by suction filtration, the crude product thus obtained was isolated by preparative HPLC to obtain 7.8 mg of the product. MS m/e 1896.5, 1893.5, 1894.5 (M+1)
(54) Step 2:
(55) ##STR00083##
(56) 5 mg of the product obtained from the previous step was dissolved in a solvent mixture of 1 ml of tetrahydrofurane and 1 ml of water, 2.0 mg of lithium hydroxide was added with stirring, the reaction solution was stirred for 1 hour, 10 mg of acetic acid was added to quench the reaction, the organic solvent was removed by a rotary evaporator, purification by preparative HPLC obtained 1.8 mg of the product, MS m/e 1881.5, 1880.5, 1879.5 (M+1)
Example 8
(57) Compounds V61, V62, V63, V64, V65, V66, V69 and the like were prepared according to the process as described in Example 7.