NOVEL ANTI-INFECTIVE AND ANTI-INFLAMMATORY COMPOUNDS

Abstract

Lysosomally accumulated substances that release a nitroxy group, or a short chain fatty acid or a product of anaerobic metabolism or a thiol or a sulfide often from an ester or similar labile linkage have anti-inflammatory, anti-cancer and anti-bacterial activity. They are useful in treating infectious, inflammatory and malignant disease.

Claims

1. A compound, or salt thereof, comprising an amphiphilic lysosomally trapped compound (alc) conjugated via an ester, thioester, amide, or nitroester to one or more products of anaerobic metabolism (pams) of the same or different types.

2. The compound as in claim 1 in which the PAM is selected from one or more of Short Chain Fatty Acid (SCFA), NO, H.sub.2S, mercaptans, polyamines, decarboxylated amino acids, TLR ligands, or polyphenol metabolites like phenylpropionic acid.

3. The compound as in claim 1 in which the ALC is selected from a macrolide, polyamine, propranolol analog, chloroquine analog, amodiaquine, dextromethorphan, dextrorphan, paroxetine, fluoxetine, astemizole or imipramine analog.

4. A method of stimulating immune or epithelial cells to form an anti-infective barrier or anti-infective response comprising contacting the cells with a compound of Formula 1, or salt thereof, wherein the contacting results in the intracellular release of a PAM comprising one or more of a molecule type selected from TLR ligands, SCFA, NO, H.sub.2S, sulfides, polyamines, decarboxylated amino acids or polyphenol metabolites like phenylpropionic acid from the compound of Formula 1, or salt thereof.

5. The method as in claim 4 comprising the intracellular release of one or more types of a short chain fatty acid moiety from the compound of Formula 1.

6. The method as in claim 5 comprising the intracellular release of a short chain fatty acid moiety containing 2 or more carbons from an appropriate carrier molecule.

7. (canceled)

8. The compound of claim 1, wherein the compound is one of the following: (Formula 2) ##STR00109## Wherein, X=—N(CH.sub.3)—CH.sub.2—; —CH.sub.2—N[(CH.sub.2).sub.n—CH.sub.3]— wherein n is 0-4; —C(═O)—; —C(═NOR.sup.8)—; —C(═NR.sup.12)—; R.sub.1 is: -(C.sub.1-C.sub.10)alkyl; -(C.sub.1-C.sub.10)alkyliden-OH; -(C.sub.1-C.sub.10)alkyliden-ONO.sub.2; R.sub.2 is: —H; —NO.sub.(y) with y=1 or 2; —C(═O)OR.sup.7, —C(═S)OR.sup.7, —C(═O)R.sup.7, —C(═S)R.sup.7, —C(═O)(NH)R.sup.7, —C(═S)(NH)R.sup.7; R.sup.3 is: —H; —NO.sub.(y) with y=1 or 2; —C(═O)OR.sup.7, —C(═S)OR.sup.7, —C(═O)R.sup.7, —C(═S)R.sup.7, —C(═O)(NH)R.sup.7, —C(═S)(NH)R.sup.7; If Z=O, R.sub.4 is: —H; —NO.sub.(y) with y=1 or 2; —C(═O)OR.sup.7, —C(═S)OR.sup.7, —C(═O)R.sup.7, —C(═S)R.sup.7, —C(═O)(NH)R.sup.7, —C(═S)(NH)R.sup.7; R.sup.5 is: —H; —NO.sub.(y) with y=1 or 2; —C(═O)OR.sup.7, —C(═S)OR.sup.7, —C(═O)R.sup.7, —C(═S)R.sup.7, —C(═O)(NH)R.sup.7, —C(═S)(NH)R.sup.7; or Z=O or NR.sup.9 and the R.sub.4 and R.sup.5 bearing atoms are connected via —C(═O)— (If Z=O: carbonate linkage, if Z=NR.sup.9: carbamate linkage); or the R.sub.4 and R.sup.5 bearing atoms are connected via W; W is: —(—)CH-(C.sub.1-C.sub.12)alkyl; —(—)CH-(C.sub.3-C.sub.12)alkenyl; —(—)CH-(C.sub.3-C.sub.12)alkynyl; —(—)CH-(C.sub.1-C.sub.8)[(C.sub.1-C.sub.4)alkoxy]alkyl; —(—)CH-(C.sub.1-C.sub.8)[(C.sub.1-C.sub.4)alkoxy]alkenyl; wherein alkyl, alkenyl, alkynyl are optionally substituted by one to five substituents selected independently from halogen, (C.sub.1-C.sub.4)alkyl, (C.sub.1-C.sub.4)alkenyl, (C.sub.1-C.sub.4)alkynyl, (C.sub.3-C.sub.7)cycloalkyl, (C.sub.1-C.sub.6)heterocycloalkyl, (C.sub.6-C.sub.10)aryl, (C.sub.1-C.sub.9)heteroaryl, (C.sub.1-C.sub.4)alkoxy, hydroxy, nitro, cyano, azido, mercapto, —NR.sup.14R.sup.15, R.sup.14C(═O)—, R.sup.14C(═O)O—R.sup.14OC(═O)O—, R.sup.14NHC(═O)—, R.sup.14C(═O)NH—, R.sup.14R.sup.15NC(═O)—, R.sup.14OC(═O)—, and —xNO.sub.2 with x=O;S;N; R.sup.6 is: —H; —NO.sub.(y) with y=1 or 2; —C(═O)OR.sup.7, —C(═S)OR.sup.7, —C(═O)R.sup.7, —C(═S)R.sup.7, —C(═O)(NH)R.sup.7, —C(═S)(NH)R.sup.7; each R.sup.7 is independently: —H; -ferrocene; -C.sub.1-C.sub.10 alkyl, alkenyl, alkynyl, aryl, heteroaryl, alkylaryl, alkylheteroaryl wherein alkyl, alkenyl, alkynyl, aryl, heteroaryl, alkylaryl and alkylheteroaryl groups are optionally substituted by one to five substituents selected independently from: ferrocene, halogen (as can be F, Cl, Br, I), (C.sub.1-C.sub.4)alkyl, (C.sub.1-C.sub.4)alkenyl, (C.sub.1-C.sub.4)alkynyl, (C.sub.3-C.sub.7)cycloalkyl, (C.sub.1-C.sub.6)heterocycloalkyl, (C.sub.6-C.sub.10)aryl, (C.sub.1-C.sub.9)heteroaryl, (C.sub.1-C.sub.4)alkoxy, hydroxyl (—OH), nitro (—NO.sub.2), cyano (—CN), azido (—N.sub.3), mercapto (—SH), —NR.sup.14R.sup.15, R.sup.14C(═O)—, R.sup.14C(═O)O—, R.sup.14OC(═O)O—, R.sup.14NHC(═O)—, R.sup.14C(═O)NH—, R.sup.14R.sup.15NC(═O)—, R.sup.14OC(═O)—, and —XNO.sub.(y) with X=O; S; N and y=1 or 2; R.sup.8 is: —H; —NO.sub.(y) with y=1 or 2; —C(═O)—R.sup.7; -(C.sub.1-C.sub.12)alkyl; -(C.sub.1-C.sub.12)alkenyl; -(C.sub.1-C.sub.12)alkynyl; -(C.sub.1-C.sub.8)[(C.sub.1-C.sub.4)alkoxy]alkyl; -(C.sub.1-C.sub.8)[(C.sub.1-C.sub.4)alkoxy]alkenyl; -(C.sub.6-C.sub.10)aryl-(C.sub.1-C.sub.5)alkyl; -(C.sub.2-C.sub.9)heteroaryl-(C.sub.1-C.sub.5)alkyl; R.sup.9 is: —H; —NO.sub.(y) with y=1 or 2; —C(═O)—R.sup.7; -(C.sub.1-C.sub.12)alkyl; -(C.sub.1-C.sub.12)alkenyl; -(C.sub.1-C.sub.12)alkynyl; -(C.sub.1-C.sub.8)[(C.sub.1-C.sub.4)alkoxy]alkyl; -(C.sub.1-C.sub.8)[(C.sub.1-C.sub.4)alkoxy]alkenyl; -(C.sub.6-C.sub.10)aryl-(C.sub.1-C.sub.5)alkyl; -(C.sub.2-C.sub.9)heteroaryl-(C.sub.1-C.sub.5)alkyl; R.sup.12 is: —H; —NO.sub.(y) with y=1 or 2; —C(═O)—R.sup.7; -(C.sub.1-C.sub.12)alkyl; -(C.sub.1-C.sub.12)alkenyl; -(C.sub.1-C.sub.12)alkynyl; -(C.sub.1-C.sub.8)[(C.sub.1-C.sub.4)alkoxy]alkyl; -(C.sub.1-C.sub.8)[(C.sub.1-C.sub.4)alkoxy]alkenyl; -(C.sub.6-C.sub.10)aryl-(C.sub.1-C.sub.5)alkyl; -(C.sub.2-C.sub.9)heteroaryl-(C.sub.1-C.sub.5)alkyl; R.sup.14, R.sup.15 are each independently: —H; -(C.sub.1-C.sub.12)alkyl; -(C.sub.1-C.sub.12)alkenyl; -(C.sub.1-C.sub.12)alkynyl; -(C.sub.1-C.sub.8)[(C.sub.1-C.sub.4)alkoxy]alkyl; -(C.sub.1-C.sub.8)[(C.sub.1-C.sub.4)alkoxy]alkenyl; -(C.sub.6-C.sub.10)aryl-(C.sub.1-C.sub.5)alkyl; -(C.sub.2-C.sub.9)heteroaryl-(C.sub.1-C.sub.5)alkyl; wherein alkyl, alkenyl, alkynyl, aryl and heteroaryl are optionally substituted by one to five substituents selected independently from halogen (as can be F, Cl, Br, I), (C.sub.1-C.sub.4)alkyl, (C.sub.1-C.sub.4)alkenyl, (C.sub.1-C.sub.4)alkynyl, (C.sub.3-C.sub.7)cycloalkyl, (C.sub.1-C.sub.6)heterocycloalkyl, (C.sub.6-C.sub.10)aryl, (C.sub.1-C.sub.9)heteroaryl, (C.sub.1-C.sub.4)alkoxy, hydroxyl (—OH), nitro (—NO.sub.2), cyano (—CN), azido (—N.sub.3), mercapto (—SH), and —XNO.sub.y with X=O; S; N and y=1 or 2; or N(R.sup.14R.sup.15) is an aziridine, azetidine, pyrrolidine, piperidine, azepane or azocane, 1-substituted piperazine, or morpholine moiety.

9. The compound of claim 1, wherein the compound is one of the following: 1. Formula 3: ##STR00110## Wherein R.sup.1,R.sup.2,R.sup.4,R.sup.5,R.sup.6, X and Z are defined as in formula 2; R.sup.3a, R.sup.3b=both —H; or if R.sup.3a is —H, then R.sup.3b is: —OH; —OR.sup.14; NR.sup.14R.sup.15; —C(═O)—R.sup.7; or R.sup.3a=R.sup.3b=(═O); =a cyclic or non-cyclic acetal; (=NR.sup.12); =a cyclic or non-cyclic aminal; —OC(═O)R.sup.7; OR.sup.14; and R.sup.7, R.sup.14, and R.sup.15 are defined as in formula 2 in claim 8; or salt thereof.

10. A compound as in: ##STR00111## Where X is O or S; When X=O, R.sub.1 is —(C═O)CH.sub.3, —(C═O)CH.sub.2CH.sub.3, —(C═O)CH.sub.2CH.sub.2CH.sub.3, —(C═O)CH.sub.2CH.sub.2COOH, —(C═O)(C═O)CH.sub.3, —(C═O)CHCHCOOH, —(C═O)CH(OH)CH.sub.3, —(C═O)C(CH.sub.3).sub.2, —(C═O)CH.sub.2CH.sub.2CH.sub.2CH.sub.3, —(C═O)CH.sub.2C(CH.sub.3).sub.2, —(C═O)CH.sub.2CH.sub.2CH.sub.2CH.sub.2Y, or —(C═O)CH(ONO.sub.2)CH.sub.3; R.sub.2=R.sub.3=H; Y=a 5-membered saturated ring containing a disulfide bond; or When X=O, R.sub.1 is —(C═O)CH.sub.3, —(C═O)CH.sub.2CH.sub.3, —(C═O)CH.sub.2CH.sub.2CH.sub.3, —(C═O)CH.sub.2CH.sub.2COOH, —(C═O)(C═O)CH.sub.3, —(C═O)CHCHCOOH, —(C═O)CH(OH)CH.sub.3, —(C═O)C(CH.sub.3).sub.2, —(C═O)CH.sub.2CH.sub.2CH.sub.2CH.sub.3, —(C═O)CH.sub.2C(CH.sub.3).sub.2, —(C═O)CH.sub.2CH.sub.2CH.sub.2CH.sub.2Y, or —(C═O)CH(ONO.sub.2)CH.sub.3; R.sub.2=CH.sub.3; R3=H; or When X=O, R.sub.1=NO.sub.2; R.sub.2 consists of linker —CH.sub.2CH.sub.2OR.sub.4, where R.sub.4 is —(C═O)CH.sub.3, —(C═O)CH.sub.2CH.sub.3, —(C═O)CH.sub.2CH.sub.2CH.sub.3, —(C═O)CH.sub.2CH.sub.2COOH, —(C═O)(C═O)CH.sub.3, —(C═O)CHCHCOOH, —(C═O)CH(OH)CH.sub.3, —(C═O)C(CH.sub.3).sub.2, —(C═O)CH.sub.2CH.sub.2CH.sub.2CH.sub.3, —(C═O)CH.sub.2C(CH.sub.3).sub.2, or —(C═O)CH.sub.2CH.sub.2CH.sub.2CH.sub.2Y; R.sub.3=H; Y =a 5-membered saturated ring containing a disulfide bond; or When X=O, R.sub.1 is —(C═O)CH.sub.3, —(C═O)CH.sub.2CH.sub.3, —(C═O)CH.sub.2CH.sub.2CH.sub.3, —(C═O)CH.sub.2CH.sub.2COOH, —(C═O)(C═O)CH.sub.3, —(C═O)CHCHCOOH, —(C═O)CH(OH)CH.sub.3, —(C═O)C(CH.sub.3).sub.2, —(C═O)CH.sub.2CH.sub.2CH.sub.2CH.sub.3, —(C═O)CH.sub.2C(CH.sub.3).sub.2, —(C═O)CH.sub.2CH.sub.2CH.sub.2CH.sub.2Y, or —(C═O)CH(ONO.sub.2)CH.sub.3; Y=a 5-membered saturated ring containing a disulfide bond; R.sub.2 consists of linker —CH.sub.2CH.sub.2OR.sub.4, where R.sub.4=NO.sub.2; R.sub.3=H; or When X=O, R.sub.1 is NO.sub.2, R.sub.2=H or CH.sub.3, R.sup.3=OR.sub.5, where R.sub.5 is —(C═O)CH.sub.3, —(C═O)CH.sub.2CH.sub.3, —(C═O)CH.sub.2CH.sub.2CH.sub.3, —(C═O)CH.sub.2CH.sub.2COOH, —(C═O)(C═O)CH.sub.3, —(C═O)CHCHCOOH, —(C═O)CH(OH)CH.sub.3, —(C═O)C(CH.sub.3).sub.2, —(C═O)CH.sub.2CH.sub.2CH.sub.2CH.sub.3, —(C═O)CH.sub.2C(CH.sub.3).sub.2, —(C═O)CH.sub.2CH.sub.2CH.sub.2CH.sub.2Y, or —(C═O)CH(ONO.sub.2)CH.sub.3; Y=a 5-membered saturated ring containing a disulfide bond; or When X=S, R.sub.1 is —(C═O)CH.sub.3, a metal salt, or forms a disulfide bridge with itself, R.sub.2=R.sub.3=H.

11. The compound of claim 3, wherein the ALC is selected from a compound of Formula 5: ##STR00112## Wherein Mac=a macrolide ring or macrolide ring system, for example, but not limited to azithromycin or gamithromycin, each without the desosamin residue.

12. The compound of claim 3, wherein the ALC is selected from a compound of Formula 6: ##STR00113## Wherein Mac=a macrolide ring or macrolide ring system, for example, but not limited to azithromycin or gamithromycin, each without the desosamin residue R′=independently of each other —H; —NO.sub.(y) with y=1 or 2; —C(═O)OR.sup.3, —C(═S)OR.sup.3, —C(═O)R.sup.3, —C(═S)R.sup.3, —C(═O)(NH)R.sup.3, —C(═S)(NH)R.sup.3; R.sup.1, R.sub.2=independently of each other H, OH, OR.sub.4, -C.sub.1-C.sub.10 alkyl, alkenyl, alkynyl, aryl, heteroaryl, alkylaryl, alkylheteroaryl; wherein alkyl, alkenyl, alkynyl, aryl, heteroaryl, alkylaryl and alkylheteroaryl groups are optionally substituted by one to five substituents selected independently from: fluorine, (C.sub.1-C.sub.4)alkyl, (C.sub.1-C.sub.4)alkenyl, (C.sub.1-C.sub.4)alkynyl, (C.sub.3-C.sub.7)cycloalkyl, (C.sub.1-C.sub.9)heterocycloalkyl, (C.sub.6-C.sub.10)aryl, (C.sub.1-C.sub.9)heteroaryl, (C.sub.1-C.sub.4)alkoxy, hydroxyl (—OH), nitro (—NO.sub.2), cyano (—CN), azido (—N.sub.3), mercapto (—SH), (C.sub.1-C.sub.4)alkthio, —NR.sup.4R.sup.5, R.sup.4C(═O)—, R.sup.4C(═O)O—, R.sup.4OC(═O)O—, R.sup.4NHC(═O)—, R.sup.4C(═O)NH—, R.sup.4R.sup.5NC(═O)—, R.sup.4OC(═O)—, and —XNO.sub.(y) with X=O; S; N and y=1 or 2; or N(R.sup.1R.sup.2) is an aziridine, azetidine, pyrrolidine, piperidine, azepane or azocane, 1-substituted piperazine, or morpholine moiety; R.sup.3=-C.sub.1-C.sub.10 alkyl, alkenyl, alkynyl, aryl, heteroaryl, alkylaryl, alkylheteroaryl wherein alkyl, alkenyl, alkynyl, aryl, heteroaryl, alkylaryl and alkylheteroaryl groups are optionally substituted by one to five substituents selected independently from: halogen, (C.sub.1-C.sub.4)alkyl, (C.sub.1-C.sub.4)alkenyl, (C.sub.1-C.sub.4)alkynyl, (C.sub.3-C.sub.7)cycloalkyl, (C.sub.1-C.sub.9)heterocycloalkyl, (C.sub.6-C.sub.10)aryl, (C.sub.1-C.sub.9)heteroaryl, (C.sub.1-C.sub.4)alkoxy, hydroxyl (—OH), nitro (—NO.sub.2), cyano (—CN), azido (—N.sub.3), mercapto (—SH), (C.sub.1-C.sub.4)alkthio, —NR.sup.6R.sup.7, R.sup.6C(═O)—, R.sup.6C(═O)O—, R.sup.6OC(═O)O—, R.sup.6NHC(═O)—, R.sup.6C(═O)NH—, R.sup.6R.sup.7NC(═O)—, R.sup.6OC(═O)—, and —XNO.sub.(y) with X=O; S; N and y=1 or 2; R.sup.4, R.sup.5, R.sup.6 and R.sup.7 are each independently: —H; -(C.sub.1-C.sub.12)alkyl; -(C.sub.1-C.sub.12)alkenyl; -(C.sub.1-C.sub.12)akynyl; -(C.sub.1-C.sub.8)[(C.sub.1-C.sub.4)alkoxy]alkyl; -(C.sub.1-C.sub.8)[(C.sub.1-C.sub.4)alkoxy]alkenyl; -(C.sub.6-C.sub.10)aryl-(C.sub.1-C.sub.5)alkyl; -(C.sub.2-C.sub.9)heteroaryl-(C.sub.1-C.sub.5)alkyl; wherein alkyl, alkenyl, alkynyl, aryl and heteroaryl are optionally substituted by one to five substituents selected independently from ferrocene, halogen, (C.sub.1-C.sub.4)alkyl, (C.sub.1-C.sub.4)alkenyl, (C.sub.1-C.sub.4)alkynyl, (C.sub.3-C.sub.7)cycloalkyl, (C.sub.1-C.sub.9)heterocycloalkyl, (C.sub.6-C.sub.10)aryl, (C.sub.1-C.sub.9)heteroaryl, (C.sub.1-C.sub.4)alkoxy, hydroxyl (—OH), (C.sub.1-C.sub.6)acyloxy, nitro (—NO.sub.2), cyano (—CN), azido (—N.sub.3), mercapto (—SH), and —XNO.sub.y with X=O; S; N and y=1 or 2.

13. The compound of claim 8, wherein the number of PAMs is 0.

14. The compound of claim 9, wherein the number of PAMs is 0.

15. A non-nitrate salt of a compound as in claim 1.

16. A pharmaceutical composition comprising a compound of claim 1, or salt, solvate, or hydrate thereof, and a pharmaceutically acceptable carrier.

17. The composition of claim 16, further comprising an additional therapeutic agent.

18. A method of treating a disease, disorder, or symptom thereof in a subject comprising administering to the subject a compound of claim 1, or salt, solvate, or hydrate thereof.

19. The method of claim 18, wherein the disease or disorder is an infectious disease, an inflammatory disease, or a malignant disease.

20. The method of claim 18, further comprising administering an antibacterial compound.

21. The method of claim 18, further comprising administering a macrolide compound.

Description

BRIEF DESCRIPTION OF DRAWINGS

[0301] FIG. 1: TNFa-Production by LPS treated mice with either Vehicle (1% citric acid in water, 5 mL/kg) or 10 μmol/kg Compound E2 or Compound E3.

[0302] FIG. 2: Change in body weight in mice in which arthritis has been induced using bovine collagen. Animals were treated with either Vehicle (1% citric acid in water, 5 mL/kg) or the indicated doses of compound E2 μmol/kg. Data are from 10 animals per group, and data points significant different from Vehicle are marked with *.

[0303] FIG. 3: Number of mice maintaining body weight in which intestinal inflammation has been induced using Dextran Sulfate. Mice were treated with Vehicle (1% citric acid in water, 5 mL/kg) or the indicated doses of compound E2 in μmol/kg.

[0304] FIG. 4: Killing of phagocytosed Salmonella typhimurium by murine macrophages treated with either the commercial antibiotic azithromycin or Compound E2. Compound E2 stimulates the killing of bacterial cells by macrophages.

[0305] FIG. 5: Effect of substance E5 on the response of mice to an infection by Staphylococcus. Treatment with the substance E5 results in a faster recovery of weight due to faster clearance of bacteria

[0306] FIG. 6: Effect of substances E2, E5 and Azithromycin on the ability of mice to clear an infection by Staphylococcus aureus Newman. Bacteria are quantified as CFU recovered from a standard sample of kidney. Treatment with substances decreases recovered bacteria in a dose responsive manner.

[0307] FIG. 7: Effect of substances on the ability of mice to tolerate dextran sulfate colitis. Cyclosporine is provided at a dose of 25 mg/kg, all other substances including azithromycin are provided at a dose of 0.1 μmol/kg.

[0308] FIG. 8: Effect of substances E5 and Cyclosporin on the liver weight of mice treated with dextran sulfate colitis. Data are the mean of N=8 and are plotted with the 95% confidence interval.

[0309] FIG. 9: Effect of substances E-5 and E-241 versus Vehicle on the development of EAE in the C57B6 mouse. Plotted is the clinical score based with antigen injected on day 0 and substance started on day 7. Data are the mean of N=8.

[0310] FIG. 10: Effect of substances compared with the positive control Cyclosporin on the body weight of mice treated with dextran sulfate colitis. Data are the mean of N=8 and are plotted with the 95% confidence interval. Above the bars are the p values vs. Vehicle for a T-test.

[0311] FIG. 11: Effect of substances compared with the positive control Cyclosporin on the colon length of mice treated with dextran sulfate colitis. Data are the mean of N=8 and are plotted with the 95% confidence interval. Above the bars are the p values vs. Vehicle for a T-test.

[0312] FIG. 12: Effect of substances compared with the positive control Cyclosporin on the amount of fluorocein labelled dextran (FITC) taken up into the serums of mice treated with dextran sulfate colitis. 4 h prior to sampling, mice are treated with an oral suspension of FITC dextran which would normally not enter the blood stream. The effect of DSS is to disrupt the gut epithelium allowing larger molecules to enter the blood stream. Reductions in FITC dextran suggest improved barrier function. Data are the mean of N=8 and are plotted with the 95% confidence interval. Above the bars are the p values vs. Vehicle for a T-test.

[0313] FIG. 13: Effect of substances compared with the positive control Cyclosporin on the serum Calcium of mice treated with dextran sulfate to induce colitis at day 8 after starting DSS. Data are the mean of N=8 and are plotted with the 95% confidence interval. Above the bars are the p values vs. Vehicle for a T-test.

[0314] FIG. 14: Effect of substances compared with the positive control Cyclosporin on the clinical score of mice treated with dextran sulfate to induce colitis at day 8 after starting DSS. Data are the mean of N=8 and are plotted with the 95% confidence interval. Above the bars are the p values vs. Vehicle for a T-test.

[0315] FIG. 15: Effect of substances compared with the positive control Cyclosporin on the serum Potassium of mice treated with dextran sulfate to induce colitis at day 8 after starting DSS. Data are the mean of N=8 and are plotted with the 95% confidence interval. Above the bars are the p values vs. Vehicle for a T-test.

[0316] FIG. 16: Effect of substances compared with the positive control Cyclosporin on the serum Total Bilirubin of mice treated with dextran sulfate to induce colitis at day 8 after starting DSS. Data are the mean of N=8 and are plotted with the 95% confidence interval. Above the bars are the p values vs. Vehicle for a T-test.

[0317] FIG. 17: Body weight of BALBc mice at day 7 after commencing 2.5% DSS in water. DSS causes lesions in the colon that lead to weight loss. Substance E-2, amongst others, protects against weight loss. Data are the mean of N=8 and are plotted with the 95% confidence interval.

[0318] FIG. 18: Body weight and clinical score of BALBc mice at day 9 after commencing 2.5% DSS in water. DSS causes lesions in the colon that lead to weight loss. Substances E-3, E-238 and E-553, amongst others, stimulate inflammation. All doses 1.34 μmol/kg. Data are the mean of N=8 and are plotted with the 95% confidence interval. Above the bars are the p values vs. Vehicle for a T-test.

[0319] FIG. 19: Body weight and clinical score of BALBc mice at day 7 after commencing 2.5% DSS in water. DSS causes lesions in the colon that lead to weight loss. Substance like E-51 with weight greater than Vehicle protect against inflammation. Doses are as indicated. Data are the mean of N=8 and are plotted with the 95% confidence interval. Above the bars are the p values vs. Vehicle for a T-test.

[0320] FIG. 20: The compounds containing R.sub.1 nitrate ester have preferential distribution to the lung. Data show the concentration of the substance in the lung and liver at 6 h after administration of a 10 mg/kg dose p.o. in 2% citric acid.

[0321] FIG. 21: The effect of various compounds on the rate of killing of Salmonella typhimurium following incubation and phagocytosis by J774 murine cells. The number of surviving bacteria is an indicator of the degree of intracellular killing of the bacteria, All substances are supplied at an initial concentration of 1 μM.

DESCRIPTION OF EMBODIMENTS

[0322] General Procedure for the Introduction of the Nitrooxide Group

[0323] The compound to be nitrated (1 equiv.) (—SH, —OH, —NH) is dissolved or suspended in acetic acid (approximately 6.0 ml per 1 mmol compound to be nitrated) and a solution of nitric acid (10% in acetic anhydride, about 3.25 ml per 1 mmol compound to be nitrated) is slowly added to the system while cooling in an ice bath. When TLC indicated complete consumption of starting materials the mixture is poured onto ice hydrolyzing any remains of acetic anhydride, followed by cautious neutralization of acid species with sodium bicarbonate. Extraction of the aqueous system with dichloromethane (3×), drying of combined organic phases over sodium sulfate and subsequent purification of crude products by column chromatography (acetone-cyclohexane 1:3.fwdarw.1:1) yields products as amorphous white foams.

[0324] The invention will be further described in the following intermediates and examples. It should be understood that these examples are for illustrative purposes only and are not to be construed as limiting this invention in any, manner.

EXAMPLES

[0325] Unless otherwise specified, all commercially available reagents and solvents were used without prior purification. All chemical structures and names are generated from ChemDraw Ultra (Cambridge).

TABLE-US-00002 TABLE 1 Examples of ALC Core Entry ALC Core Structure A-1 Azithromycin (R.sub.1 to R.sub.5 = H, R.sub.6 = CH.sub.3) [00008] A-2 Erythromycin (X = OH) (R.sub.1 to R.sub.5 = H, R.sub.6 = CH.sub.3) or Erythromycin N-Oxime (X = N—OH) [00009] A-3 Hydroxychloroquine (R.sub.1 = H, R.sub.2 = CH.sub.3) [00010] A-4 N-ethanol HCQ (R.sub.1 = H, R.sub.2 = H) [00011] A-5 Propranolol (R.sub.1 = H, R.sub.2 = CH.sub.3) [00012] A-6 N-ethanol-propranolol (R.sub.1 = H, R.sub.2 = H) [00013] A-7 4-hydroxypropranolol (R.sub.1 = H, R.sub.2 = H, R.sub.3 = H) [00014] A-8 2-(4-fluorophenyl)-3-amino-4-(3-[2,3- di{butyroyloxy}propyloxy]phenyl)- carbonylpyrazole (R.sub.1 = H, R.sub.2 = H) [00015] A-9 2-(4-pyridyl)-3-amino-4-(3-[2,3- di{butyroyloxy}propyloxy]- phenyl)carbonylpyrazole (R.sub.1 = H, R.sub.2 = H) [00016] A-10 C5Y0073 (R.sub.1 to R.sub.5 = H, R.sub.6 = CH.sub.3) or 14-(4-Dimethylamino-3-hydroxy-6- methyl-tetrahydro-pyran-2-yloxy)-5-ethyl- 1,6,7-trihydroxy-2,6,8,9,11,13,15- heptamethyl-4,16-dioxa-9-aza- bicyclo[11.2.1]hexadecan-3-one [00017] A-11/ E-16 CSY0041 (R.sub.1 to R.sub.6 = H) or 2-Ethyl-3,4,10-trihydroxy-13-(5-hydroxy- 4-methoxy-4,6-dimethyl-tetrahydro- pyran-2-yloxy)-11-(3-hydroxy-6-methyl-4- methylamino-tetrahydro-pyran-2-yloxy)- 3,5,6,8,10,12,14-heptamethyl-1-oxa-6- aza-cyclopentadecan-15-one [00018] A-12 C5Y1239 (R.sub.1 to R.sub.5 = H) or 11-(4-Dimethylamino-3-hydroxy-6- methyl-tetrahydro-pyran-2-yloxy)-2-ethyl- 3,4,10,13-tetrahydroxy-3,5, 6,8,10,12,14-heptamethyl-1-oxa-6-aza- cyclopentadecan-15-one [00019] A-13.1 CSY1130 (R.sub.1 to R.sub.6 = H) (preparation see Example or 11-{4-[Bis-(2-hydroxy-ethyl)-amino]-3- hydroxy-6-methyl-tetrahydro-pyran-2- yloxy}-2-ethyl-3,4,10-trihydroxy-13-(5- hydroxy-4-methoxy-4,6-dimethyl- tetrahydro-pyran-2-yloxy)- 3,5,6,8,10,12,14-heptamethyl-1- oxa-6-aza-cyclopentadecan-15-one [00020] A-13.2 CSY5632 (R.sub.1 to R.sub.6 = H) or (2R,3S,4R,5R,8R,10R,11R,12S,13S,14R)- 11-(((2S,3S,6R)-3-(bis(2- hydroxyethyl)amino)-4-hydroxy-6- methyltetrahydro-2H-pyran-2-yl)oxy)-2- ethyl-3,4,10-trihydroxy-13- (((2R,4R,5S,6S)-5-hydroxy-4-methoxy-4,6- dimethyltetrahydro-2H-pyran-2-yl)oxy)- 3,5,6,8,10,12,14-heptamethyl-1-oxa-6- azacydopentadecan-15-one [00021] A-14.1 CSY2219 (R.sub.1 to R.sub.5 = H) or 2-Ethyl-3,4,10-trihydroxy-13-(5-hydroxy- 4-methoxy-4,6-dimethyl-tetrahydro- pyran-2-yloxy)-11-(3-hydroxy-6-methyl-4- morpholin-4-yl-tetrahydro-pyran-2- yloxy)-3,5,6,8,10,12,14-heptamethyl-1- oxa-6-aza-cyclopentadecan-15-one [00022] A-14.2 CSY5602 (R.sub.1 to R.sub.5 = H) or (2R,3S,4R,5R,8R,10R,11R,12S,13S,14R)-2- ethyl-3,4,10-trihydroxy-13- (((2R,4R,5S,6S)-5-hydroxy-4-methoxy-4,6- dimethyltetrahydro-2H-pyran-2-yl)oxy)- 11-(((2S,3S,6R)-4-hydroxy-6-methyl-3- morpholinotetrahydro-2H-pyran-2- yl)oxy)-3,5,6,8,10,12,14-heptamethyl-1- oxa-6-azacyclopentadecan-15-one [00023] A-15 CSY1019 (R.sub.1 to R.sub.5 = H) or 11-(4-Dimethylamino-6-methyl-3- nitrooxy-tetrahydro-pyran-2-yloxy)-2- ethyl-3,4,10-trihydroxy-13-(5-hydroxy-4- methoxy-4,6-dimethyl-tetrahydro-pyran- 2-yloxy)-3,5,6,8,10,12,14-heptamethyl-1- oxa-6-aza-cyclopentadecan-15-one [00024] A-16 Tildipirosin (R.sub.1 to R.sub.3 = H) [00025] A-17 Gamithromycin (R.sub.1 to R.sub.5 = H) [00026] A-18 Tylosin (R.sub.1 to R.sub.5 = H) [00027] A-19.1 Polyamines (R.sub.1 to R.sub.2 = H) [00028] A-19.2 Polyamines (R.sub.1 to R.sub.4 = H) [00029] A-19.3 Polyamines (R.sub.1 = R.sub.2 = H; R.sub.3 = alkyl, usually ethyl) [00030] A-20.1 Tris(hydroxymethyl)nitromethane (R.sub.1 to R.sub.3 = H) [00031] A-20.2 Sodium Tris(hydroxymethyl)aminopropylsulfonate (R.sub.1 to R.sub.3 = H) [00032] A-21 Clarithromycin (R.sub.1 to R.sub.4 = H) [00033] A-22 Tulathromycin (R.sub.1 to R.sub.5 = 0) [00034]

TABLE-US-00003 TABLE 2 Examples of compounds showing the appropriate substituents. Compound formulae based on “ALC core” structures as detailed in Table 1 (above). Compound Entry ALC Core R.sub.1 R.sub.2 R.sub.3 R.sub.4 R.sub.5 R.sub.6 E-1 Azithromycin NO.sub.2 H H H H CH.sub.3 E-2 Azithromycin NO.sub.2 Ac H H H CH.sub.3 E-3 Azithromycin NO.sub.2 H H H H C-10 alkyl E-4 Azithromycin NO.sub.2 Ac H H H C-10 alkyl E-5 Azithromycin NO.sub.2 Propionyl H H H CH.sub.3 E-6 Azithromycin p-Nitro NO.sub.2 NO.sub.2 H H CH.sub.3 benzoyl E-7 Azithromycin Benzoyl NO.sub.2 H H H CH.sub.3 E-8 Erythromycin NO.sub.2 Ac H H H CH.sub.3 oxime E-9 Azithromycin Ac NO.sub.2 H H H CH.sub.3 E-10 Azithromycin H NO.sub.2 H H H CH.sub.3 E-11 Azithromycin NO.sub.2 ch.sub.3 H H H CH.sub.3 E-12 Azithromycin H Tetranitro H H H CH.sub.3 moiety.sup.1 E-13 Azithromycin NO.sub.2 Propionyl H H Propionyl CH.sub.3 E-14 Azithromycin H H H H H Propanol- NO.sub.2 E-15 Azithromycin H H H H H CH.sub.3 E-16 Azithromycin H H H H H H E-17/I-7 Azithromycin H Ac H H H CH.sub.3 E-18 Azithromycin H Propionyl H H H CH.sub.3 E-19 Azithromycin H Butyryl H H H CH.sub.3 E-20/I-2 Azithromycin H H H H H C-10 alkyl E-21 Azithromycin NO.sub.2 NO.sub.2 H H H CH.sub.3 E-22/I-4 Azithromycin Ac CH.sub.2CCH H H H CH.sub.3 E-23/I-6 Azithromycin Ac Ac H H H CH.sub.3 E-24 Azithromycin NO.sub.2 Butyryl H H H CH.sub.3 E-25//I-3 Azithromycin Ac H H H H CH.sub.3 E-26/I-8 Azithromycin Benzoyl H H H H CH.sub.3 E-27 Azithromycin Succinyl H H H H CH.sub.3 E-28 Azithromycin Ac Propionyl H H H CH.sub.3 E-29 Azithromycin Ac Butyryl H H H CH.sub.3 E-30 Azithromycin Propionyl H H H H CH.sub.3 E-31 Azithromycin Propionyl NO.sub.2 H H H CH.sub.3 E-32 Azithromycin Propionyl Ac H H H CH.sub.3 E-33 Azithromycin Propionyl Propionyl H H H CH.sub.3 E-34 Azithromycin Propionyl Butyryl H H H CH.sub.3 E-35 Azithromycin Butyryl H H H H CH.sub.3 E-36 Azithromycin Butyryl NO.sub.2 H H H CH.sub.3 E-37 Azithromycin Butyryl Ac H H H CH.sub.3 E-38 Azithromycin Butyryl Propionyl H H H CH.sub.3 E-39 Azithromycin Butyryl Butyryl H H H CH.sub.3 E-40 Azithromycin H Succinyl H H H CH.sub.3 E-41 Azithromycin H Pyruvyl H H H CH.sub.3 E-42 Azithromycin H Maleyl H H H CH.sub.3 E-43 Azithromycin H Lactyl H H H CH.sub.3 E-44 Azithromycin H Isobutyryl H H H CH.sub.3 E-45 Azithromycin H Valeryl H H H CH.sub.3 E-46 Azithromycin H Isovaleryl H H H CH.sub.3 E-47 Azithromycin Butyryl Butyryl Butyryl Butyryl H CH.sub.3 E-48 Azithromycin Butyryl Butyryl Butyryl H H CH.sub.3 E-49 Azithromycin Ac Ac Ac Ac H CH.sub.3 E-50 Azithromycin Ac Ac Ac H H CH.sub.3 E-51 Azithromycin Propionyl Propionyl Propionyl Propionyl H CH.sub.3 E-52 Azithromycin Propionyl Propionyl Propionyl H H CH.sub.3 E-53 Azithromycin Succinyl Succinyl Succinyl Succinyl H CH.sub.3 E-54 Azithromycin Pyruvyl Pyruvyl Pyruvyl Pyruvyl H CH.sub.3 E-55 Azithromycin Maleyl Maleyl Maleyl Maleyl H CH.sub.3 E-56 Azithromycin Lactyl Lactyl Lactyl Lactyl H CH.sub.3 E-57 Azithromycin Isobutyryl Isobutyryl Isobutyryl Isobutyryl H CH.sub.3 E-58 Azithromycin Valeryl Valeryl Valeryl Valeryl H CH.sub.3 E-59 Azithromycin Isovaleryl Isovaleryl Isovaleryl Isovaleryl H CH.sub.3 E-60 Azithromycin Succinyl Succinyl Succinyl H H CH.sub.3 E-61 Azithromycin Pyruvyl Pyruvyl Pyruvyl H H CH.sub.3 E-62 Azithromycin Maleyl Maleyl Maleyl H H CH.sub.3 E-63 Azithromycin Lactyl Lactyl Lactyl H H CH.sub.3 E-64 Azithromycin Isobutyryl Isobutyryl Isobutyryl H H CH.sub.3 E-65 Azithromycin Valeryl Valeryl Valeryl H H CH.sub.3 E-66 Azithromycin Isovaleryl Isovaleryl Isovaleryl H H CH.sub.3 E-67 Azithromycin Succinyl Succinyl H H H CH.sub.3 E-68 Azithromycin Pyruvyl Pyruvyl H H H CH.sub.3 E-69 Azithromycin Maleyl Maleyl H H H CH.sub.3 E-70 Azithromycin Lactyl Lactyl H H H CH.sub.3 E-71 Azithromycin Isobutyryl Isobutyryl H H H CH.sub.3 E-72 Azithromycin Valeryl Valeryl H H H CH.sub.3 E-73 Azithromycin Isovaleryl Isovaleryl H H H CH.sub.3 E-74 Azithromycin Acetoxypropionyl H H H H CH.sub.3 E-75 Azithromycin Lipoyl Lipoyl H H H CH.sub.3 E-76 Azithromycin H Lipoyl H H H CH.sub.3 E-77 Azithromycin Lipoyl H H H H CH.sub.3 E-78 Azithromycin NO.sub.2 Lipoyl H H H CH.sub.3 E-79 Azithromycin NO.sub.2 Succinyl - H H H CH.sub.3 dithiole- 3-thione E-80 Azithromycin Succinyl - H H H H CH.sub.3 dithiole- 3-thione E-81 Azithromycin Polysulfide H H H H CH.sub.3 ethyl carbonate E-82 Azithromycin NO- H H H H CH.sub.3 thioethylcarbonate E-83 Azithromycin O-Phenyl H H H H CH.sub.3 chlorothionocarbonate E-84 Azithromycin n- H H H H CH.sub.3 hexanoyl E-85 Azithromycin Bromoethylcarbonate H H H H CH.sub.3 E-86 Azithromycin Vinyll H H H H CH.sub.3 carbonate E-87 Hydroxychloroquine Ac H E-88 Hydroxychloroquine Propionyl H E-89 Hydroxychloroquine Butyryl H E-90 Hydroxychloroquine Succinyl H E-91 Hydroxychloroquine Pyruvyl H E-92 Hydroxychloroquine Maleyl H E-93 Hydroxychloroquine Lactyl H E-94 Hydroxychloroquine Isobutyryl H E-95 Hydroxychloroquine Valeryl H E-96 Hydroxychloroquine Isovaleryl H E-97 Hydroxychloroquine Lipoyl H E-98 Hydroxychloroquine Ac CH.sub.3 E-99 Hydroxychloroquine Propionyl CH.sub.3 E-100 Hydroxychloroquine Butyryl CH.sub.3 E-101 Hydroxychloroquine Succinyl CH.sub.3 E-102 Hydroxychloroquine Pyruvyl CH.sub.3 E-103 Hydroxychloroquine Maleyl CH.sub.3 E-104 Hydroxychloroquine Lactyl CH.sub.3 E-105 Hydroxychloroquine Isobutyryl CH.sub.3 E-106 Hydroxychloroquine Valeryl CH.sub.3 E-107 Hydroxychloroquine Isovaleryl CH.sub.3 E-108 Hydroxychloroquine Lipoyl CH.sub.3 E-109 N-ethanol HCQ NO.sub.2 Ac E-110 N-ethanol HCQ NO.sub.2 Propionyl E-111 N-ethanol HCQ NO.sub.2 Butyryl E-112 N-ethanol HCQ NO.sub.2 Succinyl E-113 N-ethanol HCQ NO.sub.2 Pyruvyl E-114 N-ethanol HCQ NO.sub.2 Maleyl E-115 N-ethanol HCQ NO.sub.2 Lactyl E-116 N-ethanol HCQ NO.sub.2 Isobutyryl E-117 N-ethanol HCQ NO.sub.2 Valeryl E-118 N-ethanol HCQ NO.sub.2 Isovaleryl E-119 N-ethanol HCQ NO.sub.2 Lipoyl E-120 N-ethanol HCQ Ac NO.sub.2 E-121 N-ethanol HCQ Propionyl NO.sub.2 E-122 N-ethanol HCQ Butyryl NO.sub.2 E-123 N-ethanol HCQ Succinyl NO.sub.2 E-124 N-ethanol HCQ Pyruvyl NO.sub.2 E-125 N-ethanol HCQ Maleyl NO.sub.2 E-126 N-ethanol HCQ Lactyl NO.sub.2 E-127 N-ethanol HCQ Isobutyryl NO.sub.2 E-128 N-ethanol HCQ Valeryl NO.sub.2 E-129 N-ethanol HCQ Isovaleryl NO.sub.2 E-130 N-ethanol HCQ Lipoyl NO.sub.2 E-131 Propranolol Ac H E-132 Propranolol Propionyl H E-133 Propranolol Butyryl H E-134 Propranolol Succinyl H E-135 Propranolol Pyruvyl H E-136 Propranolol Maleyl H E-137 Propranolol Lactyl H E-138 Propranolol Isobutyryl H E-139 Propranolol Valeryl H E-140 Propranolol Isovaleryl H E-141 Propranolol Lipoyl H E-142 Propranolol 2-O- H Nitrolactyl E-143 Propranolol Ac CH.sub.3 E-144 Propranolol Propionyl CH.sub.3 E-145 Propranolol Butyryl CH.sub.3 E-146 Propranolol Succinyl CH.sub.3 E-147 Propranolol Pyruvyl CH.sub.3 E-148 Propranolol Maleyl CH.sub.3 E-149 Propranolol Lactyl CH.sub.3 E-150 Propranolol Isobutyryl CH.sub.3 E-151 Propranolol Valeryl CH.sub.3 E-152 Propranolol Isovaleryl CH.sub.3 E-153 Propranolol Lipoyl CH.sub.3 E-154 N-ethanol- NO.sub.2 Ac propranolol E-155 N-ethanol- NO.sub.2 Propionyl propranolol E-156 N-ethanol- NO.sub.2 Butyryl propranolol E-157 N-ethanol- NO.sub.2 Succinyl propranolol E-158 N-ethanol- NO.sub.2 Pyruvyl propranolol E-159 N-ethanol- NO.sub.2 Maleyl propranolol E-160 N-ethanol- NO.sub.2 Lactyl propranolol E-161 N-ethanol- NO.sub.2 Isobutyryl propranolol E-162 N-ethanol- NO.sub.2 Valeryl propranolol E-163 N-ethanol- NO.sub.2 Isovaleryl propranolol E-164 N-ethanol- NO.sub.2 Lipoyl propranolol E-165 N-ethanol- Propionyl NO.sub.2 propranolol E-166 N-ethanol- Butyryl NO.sub.2 propranolol E-167 N-ethanol- Succinyl NO.sub.2 propranolol E-168 N-ethanol- Pyruvyl NO.sub.2 propranolol E-169 N-ethanol- Maleyl NO.sub.2 propranolol E-170 N-ethanol- Lactyl NO.sub.2 propranolol E-171 N-ethanol- Isobutyryl NO.sub.2 propranolol E-172 N-ethanol- Valeryl NO.sub.2 propranolol E-173 N-ethanol- Isovaleryl NO.sub.2 propranolol E-174 N-ethanol- Lipoyl NO.sub.2 propranolol E-175 4-hydroxy NO.sub.2 H Ac propranolol E-176 4-hydroxy NO.sub.2 H Propionyl propranolol E-177 4-hydroxy NO.sub.2 H Butyryl propranolol E-178 4-hydroxy NO.sub.2 H Succinyl propranolol E-179 4-hydroxy NO.sub.2 H Pyruvyl propranolol E-180 4-hydroxy NO.sub.2 H Maleyl propranolol E-181 4-hydroxy NO.sub.2 H Lactyl propranolol E-182 4-hydroxy NO.sub.2 H Isobutyryl propranolol E-183 4-hydroxy NO.sub.2 H Valeryl propranolol E-184 4-hydroxy NO.sub.2 H Isovaleryl propranolol E-185 4-hydroxy NO.sub.2 H Lipoyl propranolol E-186 4-hydroxy NO.sub.2 CH.sub.3 Ac propranolol E-187 4-hydroxy NO.sub.2 CH.sub.3 Propionyl propranolol E-188 4-hydroxy NO.sub.2 CH.sub.3 Butyryl propranolol E-189 4-hydroxy NO.sub.2 CH.sub.3 Succinyl propranolol E-190 4-hydroxy NO.sub.2 CH.sub.3 Pyruvyl propranolol E-191 4-hydroxy NO.sub.2 CH.sub.3 Maleyl propranolol E-192 4-hydroxy NO.sub.2 CH.sub.3 Lactyl propranolol E-193 4-hydroxy NO.sub.2 CH.sub.3 Isobutyryl propranolol E-194 4-hydroxy NO.sub.2 CH.sub.3 Valeryl propranolol E-195 4-hydroxy NO.sub.2 CH.sub.3 Isovaleryl propranolol E-196 4-hydroxy NO.sub.2 CH.sub.3 Lipoyl propranolol E-197 A-8 Ac Ac E-198 A-8 Propionyl Propionyl E-199 A-8 Butyryl Butyryl E-200 A-8 Succinyl Succinyl E-201 A-8 Pyruvyl Pyruvyl E-202 A-8 Maleyl Maleyl E-203 A-8 Lactyl Lactyl E-204 A-8 Isobutyryl Isobutyryl E-205 A-8 Valeryl Valeryl E-206 A-8 Isovaleryl Isovaleryl E-207 A-8 Lipoyl Lipoyl E-208 A-9 Ac Ac E-209 A-9 Propionyl Propionyl E-210 A-9 Butyryl Butyryl E-211 A-9 Succinyl Succinyl E-212 A-9 Pyruvyl Pyruvyl E-213 A-9 Maleyl Maleyl E-214 A-9 Lactyl Lactyl E-215 A-9 Isobutyryl Isobutyryl E-216 A-9 Valeryl Valeryl E-217 A-9 Isovaleryl Isovaleryl E-218 A-9 Lipoyl Lipoyl E-219 A-10 H Tetranitro H H CH.sub.3 moiety1 E-220 A-10 H H H H Propanol- NO.sub.2 E-221 A-10 H H H H CH.sub.3 E-222 A-10 H H H H H E-223 A-10 H Ac H H CH.sub.3 E-224 A-10 H Propionyl H H CH.sub.3 E-225 A-10 H Butyryl H H CH.sub.3 E-226 A-10 H H H H C-10 alkyl E-227 A-10 Ac CH.sub.2CCH H H CH.sub.3 E-228 A-10 Ac Ac H H CH.sub.3 E-229 A-10 Ac H H H CH.sub.3 E-230 A-10 Benzoyl H H H CH.sub.3 E-231 A-10 Succinyl H H H CH.sub.3 E-232 A-10 Ac Propionyl H H CH.sub.3 E-233 A-10 Ac Butyryl H H CH.sub.3 E-234 A-10 Propionyl H H H CH.sub.3 E-235 A-10 Propionyl Ac H H CH.sub.3 E-236 A-10 Propionyl Propionyl H H CH.sub.3 E-237 A-10 Propionyl Butyryl H H CH.sub.3 E-238 A-10 Butyryl H H H CH.sub.3 E-239 A-10 Butyryl Ac H H CH.sub.3 E-240 A-10 Butyryl Propionyl H H CH.sub.3 E-241 A-10 Butyryl Butyryl H H CH.sub.3 E-242 A-10 H Succinyl H H CH.sub.3 E-243 A-10 H Pyruvyl H H CH.sub.3 E-244 A-10 H Maleyl H H CH.sub.3 E-245 A-10 H Lactyl H H CH.sub.3 E-246 A-10 H Isobutyryl H H CH.sub.3 E-247 A-10 H Valeryl H H CH.sub.3 E-248 A-10 H Isovaleryl H H CH.sub.3 E-249 A-10 Butyryl Butyryl Butyryl H CH.sub.3 E-250 A-10 Butyryl Butyryl H H CH.sub.3 E-251 A-10 Ac Ac Ac H CH.sub.3 E-252 A-10 Ac Ac H H CH.sub.3 E-253 A-10 Propionyl Propionyl Propionyl H CH.sub.3 E-254 A-10 Propionyl Propionyl H H CH.sub.3 E-255 A-11/E-16 Butyric H H H H Butyric E-256 A-11/E-16 Butyric Butyric H H H Butyric E-257 A-11/E-16 Butyric Butyric Butyric H H Butyric E-258 A-11/E-16 H H H H H Mannose E-259 A-11/E-16 Propionic H H H H Propionic E-260 A-11/E-16 Propionic Propionic H H H Propionic E-261 A-11/E-16 Propionic Propionic Propionic H H Propionic E-262 A-11/E-16 Ac H H H H Ac E-263 A-11/E-16 Ac Ac H H H Ac E-264 A-11/E-16 Ac Ac Ac H H Ac E-265 A-12 NO2 H H H H E-266 A-12 Ac Ac H H H E-267 A-12 Ac Ac Ac H H E-268 A-12 Ac Ac Ac Ac H E-269 A-12 Propionic Propionic H H H E-270 A-12 Propionic Propionic Propionic H H E-271 A-12 Propionic Propionic Propionic Propionic H E-272 A-12 Butyric Butyric H H H E-273 A-12 Butyric Butyric Butyric H H E-274 A-12 Butyric Butyric Butyric Butyric H E-275 A-12 Succinic Succinic H H H E-276 A-12 Pyruvic Pyruvic H H H E-277 A-12 Maleic Maleic H H H E-278 A-12 Lactic Lactic H H H E-279 A-12 Isobutyric Isobutyric H H H E-280 A-12 Isobutyric Isobutyric Isobutyric H H E-281 A-12 Isobutyric Isobutyric Isobutyric Isobutyric H E-282 A-12 Valeric Valeric H H H E-283 A-12 Valeric Valeric Valeric H H E-284 A-12 Valeric Valeric Valeric Valeric H E-285 A-12 Isovaleric Isovaleric H H H E-286 A-12 Isovaleric Isovaleric Isovaleric H H E-287 A-12 Isovaleric Isovaleric Isovaleric Isovaleric H E-288 A-12 Lipoic Lipoic H H H E-289 A-12 Lipoic Lipoic Lipoic H H E-290 A-12 Lipoic Lipoic Lipoic Lipoic H E-291 A-13.1 H H H H H Ac E-292 A-13.1 Ac H H H H Ac E-293 A-13.1 Ac Ac H H H Ac E-294 A-13.1 Ac Ac Ac H H Ac E-295 A-13.1 H H H H H Propionic E-296 A-13.1 Propionic H H H H Propionic E-297 A-13.1 Propionic Propionic H H H Propionic E-298 A-13.1 Propionic Propionic Propionic H H Propionic E-299 A-13.1 H H H H H Butyric E-300 A-13.1 Butyric H H H H Butyric E-301 A-13.1 Butyric Butyric H H H Butyric E-302 A-13.1 Butyric Butyric Butyric H H Butyric E-303 A-13.1 H H H H H Succinic E-304 A-13.1 H H H H H Pyruvic E-305 A-13.1 H H H H H Maleic E-306 A-13.1 H H H H H Lactic E-307 A-13.1 H H H H H Isobutyric E-308 A-13.1 Isobutyric H H H H Isobutyric E-309 A-13.1 Isobutyric Isobutyric H H H Isobutyric E-310 A-13.1 Isobutyric Isobutyric Isobutyric H H Isobutyric E-311 A-13.1 H H H H H Valeric E-312 A-13.1 Valeric H H H H Valeric E-313 A-13.1 Valeric Valeric H H H Valeric E-314 A-13.1 Valeric Valeric Valeric H H Valeric E-315 A-13.1 H H H H H Isovaleric E-316 A-13.1 Isovaleric H H H H Isovaleric E-317 A-13.1 Isovaleric Isovaleric H H H Isovaleric E-318 A-13.1 Isovaleric Isovaleric Isovaleric H H Isovaleric E-319 A-13.1 H H H H H Lipoic E-320 A-13.1 Lipoic H H H H Lipoic E-321 A-13.1 Lipoic Lipoic H H H Lipoic E-322 A-13.1 Lipoic Lipoic Lipoic H H Lipoic E-323 A-13.2 H H H H H Ac E-324 A-13.2 Ac H H H H Ac E-325 A-13.2 Ac Ac H H H Ac E-326 A-13.2 Ac Ac Ac H H Ac E-327 A-13.2 H H H H H Propionic E-328 A-13.2 Propionic H H H H Propionic E-329 A-13.2 Propionic Propionic H H H Propionic E-330 A-13.2 Propionic Propionic Propionic H H Propionic E-331 A-13.2 H H H H H Butyric E-332 A-13.2 Butyric H H H H Butyric E-333 A-13.2 Butyric Butyric H H H Butyric E-334 A-13.2 Butyric Butyric Butyric H H Butyric E-335 A-13.2 H H H H H Succinic E-336 A-13.2 H H H H H Pyruvic E-337 A-13.2 H H H H H Maleic E-338 A-13.2 H H H H H Lactic E-339 A-13.2 H H H H H Isobutyric E-340 A-13.2 Isobutyric H H H H Isobutyric E-341 A-13.2 Isobutyric Isobutyric H H H Isobutyric E-342 A-13.2 Isobutyric Isobutyric Isobutyric H H Isobutyric E-343 A-13.2 H H H H H Valeric E-344 A-13.2 Valeric H H H H Valeric E-345 A-13.2 Valeric Valeric H H H Valeric E-346 A-13.2 Valeric Valeric Valeric H H Valeric E-347 A-13.2 H H H H H Isovaleric E-348 A-13.2 Isovaleric H H H H Isovaleric E-349 A-13.2 Isovaleric Isovaleric H H H Isovaleric E-350 A-13.2 Isovaleric Isovaleric Isovaleric H H Isovaleric E-351 A-13.2 H H H H H Lipoic E-352 A-13.2 Lipoic H H H H Lipoic E-353 A-13.2 Lipoic Lipoic H H H Lipoic E-354 A-13.2 Lipoic Lipoic Lipoic H H Lipoic E-355 A-14.1 Ac H H H H E-356 A-14.1 Ac Ac H H H E-357 A-14.1 Ac Ac Ac H H E-358 A-14.1 Propionic H H H H E-359 A-14.1 Propionic Propionic H H H E-360 A-14.1 Propionic Propionic Propionic H H E-361 A-14.1 Butyric H H H H E-362 A-14.1 Butyric Butyric H H H E-363 A-14.1 Butyric Butyric Butyric H H E-364 A-14.1 Succinic H H H H E-365 A-14.1 Pyruvic H H H H E-366 A-14.1 Maleic H H H H E-367 A-14.1 Lactic H H H H E-368 A-14.1 Isobutyric H H H H E-369 A-14.1 Isobutyric Isobutyric H H H E-370 A-14.1 Isobutyric Isobutyric Isobutyric H H E-371 A-14.1 Valeric H H H H E-372 A-14.1 Valeric Valeric H H H E-373 A-14.1 Valeric Valeric Valeric H H E-374 A-14.1 Isovaleric H H H H E-375 A-14.1 Isovaleric Isovaleric H H H E-376 A-14.1 Isovaleric Isovaleric Isovaleric H H E-377 A-14.1 Lipoic H H H H E-378 A-14.1 Lipoic Lipoic H H H E-379 A-14.1 Lipoic Lipoic Lipoic H H E-380 A-14.2 Ac H H H H E-381 A-14.2 Ac Ac H H H E-382 A-14.2 Ac Ac Ac H H E-383 A-14.2 Propionic H H H H E-384 A-14.2 Propionic Propionic H H H E-385 A-14.2 Propionic Propionic Propionic H H E-386 A-14.2 Butyric H H H H E-387 A-14.2 Butyric Butyric H H H E-388 A-14.2 Butyric Butyric Butyric H H E-389 A-14.2 Succinic H H H H E-390 A-14.2 Pyruvic H H H H E-391 A-14.2 Maleic H H H H E-392 A-14.2 Lactic H H H H E-393 A-14.2 Isobutyric H H H H E-394 A-14.2 Isobutyric Isobutyric H H H E-395 A-14.2 Isobutyric Isobutyric Isobutyric H H E-396 A-14.2 Valeric H H H H E-397 A-14.2 Valeric Valeric H H H E-398 A-14.2 Valeric Valeric Valeric H H E-399 A-14.2 Isovaleric H H H H E-400 A-14.2 Isovaleric Isovaleric H H H E-401 A-14.2 Isovaleric Isovaleric Isovaleric H H E-402 A-14.2 Lipoic H H H H E-403 A-14.2 Lipoic Lipoic H H H E-404 A-14.2 Lipoic Lipoic Lipoic H H E-405 A-15 Succinic H H H E-406 A-15 Pyruvic H H H E-407 A-15 Maleic H H H E-408 A-15 Lactic H H H E-409 A-15 Isobutyric H H H E-410 A-15 Isobutyric Isobutyric H H E-411 A-15 Valeric H H H E-412 A-15 Valeric Valeric H H E-413 A-15 Isovaleric H H H E-414 A-15 Isovaleric Isovaleric H H E-415 A-15 Lipoic Lipoic H H E-416 A-16 Ac H H E-417 A-16 Ac Ac H E-418 A-16 Ac Ac Ac E-419 A-16 Propionic H H E-420 A-16 Propionic Propionic H E-421 A-16 Propionic Propionic Propionic E-422 A-16 Butyric H H E-423 A-16 Butyric Butyric H E-424 A-16 Butyric Butyric Butyric E-425 A-16 Isobutyric H H E-426 A-16 Isobutyric Isobutyric H E-427 A-16 Isobutyric Isobutyric Isobutyric E-428 A-16 Valeric H H E-429 A-16 Valeric Valeric H E-430 A-16 Valeric Valeric Valeric E-431 A-16 Isovaleric H H E-432 A-16 Isovaleric Isovaleric H E-433 A-16 Isovaleric Isovaleric Isovaleric E-434 A-16 Lipoic H H E-435 A-16 Lipoic Lipoic H E-436 A-16 Lipoic Lipoic Lipoic E-437 A-16 Hexanoic H H E-438 A-16 Hexanoic Hexanoic H E-439 A-16 Hexanoic Hexanoic Hexanoic E-440 A-16 Heptanoic H H E-441 A-16 Heptanoic Heptanoic H E-442 A-16 Heptanoic Heptanoic Heptanoic E-443 A-16 Octanoic H H E-444 A-16 Octanoic Octanoic H E-445 A-16 Octanoic Octanoic Octanoic E-446 A-16 Decanoic H H E-447 A-16 Decanoic Decanoic H E-448 A-16 Decanoic Decanoic Decanoic E-449 A-16 Dodecanoic H H E-450 A-16 Dodecanoic Dodecanoic H E-451 A-16 Dodecanoic Dodecanoic Dodecanoic E-452 A-17 Ac H H H H E-453 A-17 Ac Ac H H H E-454 A-17 Ac Ac Ac H H E-455 A-17 Propionic H H H H E-456 A-17 Propionic Propionic H H H E-457 A-17 Propionic Propionic Propionic H H E-458 A-17 Butyric H H H H E-459 A-17 Butyric Butyric H H H E-460 A-17 Butyric Butyric Butyric H H E-461 A-17 Isobutyric H H H H E-462 A-17 Isobutyric Isobutyric H H H E-463 A-17 Isobutyric Isobutyric Isobutyric H H E-464 A-17 Valeric H H H H E-465 A-17 Valeric Valeric H H H E-466 A-17 Valeric Valeric Valeric H H E-467 A-17 Isovaleric H H H H E-468 A-17 Isovaleric Isovaleric H H H E-469 A-17 Isovaleric Isovaleric Isovaleric H H E-470 A-17 Lipoic H H H H E-471 A-17 Lipoic Lipoic H H H E-472 A-17 Lipoic Lipoic Lipoic H H E-473 A-17 Hexanoic H H H H E-474 A-17 Hexanoic Hexanoic H H H E-475 A-17 Hexanoic Hexanoic Hexanoic H H E-476 A-17 Heptanoic H H H H E-477 A-17 Heptanoic Heptanoic H H H E-478 A-17 Heptanoic Heptanoic Heptanoic H H E-479 A-17 Octanoic H H H H E-480 A-17 Octanoic Octanoic H H H E-481 A-17 Octanoic Octanoic Octanoic H H E-482 A-17 Decanoic H H H H E-483 A-17 Decanoic Decanoic H H H E-484 A-17 Decanoic Decanoic Decanoic H H E-485 A-17 Dodecanoic H H H H E-486 A-17 Dodecanoic Dodecanoic H H H E-487 A-17 Dodecanoic Dodecanoic Dodecanoic H H E-488 A-18 Ac H H H H E-489 A-18 Ac Ac H H H E-490 A-18 Ac Ac Ac H H E-491 A-18 Propionic H H H H E-492 A-18 Propionic Propionic H H H E-493 A-18 Propionic Propionic Propionic H H E-494 A-18 Butyric H H H H E-495 A-18 Butyric Butyric H H H E-496 A-18 Butyric Butyric Butyric H H E-497 A-18 Isobutyric H H H H E-498 A-18 Isobutyric Isobutyric H H H E-499 A-18 Isobutyric Isobutyric Isobutyric H H E-500 A-18 Valeric H H H H E-501 A-18 Valeric Valeric H H H E-502 A-18 Valeric Valeric Valeric H H E-503 A-18 Isovaleric H H H H E-504 A-18 Isovaleric Isovaleric H H H E-505 A-18 Isovaleric Isovaleric Isovaleric H H E-506 A-18 Lipoic H H H H E-507 A-18 Lipoic Lipoic H H H E-508 A-18 Lipoic Lipoic Lipoic H H E-509 A-18 Hexanoic H H H H E-510 A-18 Hexanoic Hexanoic H H H E-511 A-18 Hexanoic Hexanoic Hexanoic H H E-512 A-18 Heptanoic H H H H E-513 A-18 Heptanoic Heptanoic H H H E-514 A-18 Heptanoic Heptanoic Heptanoic H H E-515 A-18 Octanoic H H H H E-516 A-18 Octanoic Octanoic H H H E-517 A-18 Octanoic Octanoic Octanoic H H E-518 A-18 Decanoic H H H H E-519 A-18 Decanoic Decanoic H H H E-520 A-18 Decanoic Decanoic Decanoic H H E-521 A-18 Dodecanoic H H H H E-522 A-18 Dodecanoic Dodecanoic H H H E-523 A-18 Dodecanoic Dodecanoic Dodecanoic H H E-524 A-19.1 Ac Ac E-525 A-19.1 Propionic Propionic E-526 A-19.1 Butyric Butyric E-527 A-19.1 Isobutyric Isobutyric E-528 A-19.1 Valeric Valeric E-529 A-19.1 Isovaleric Isovaleric E-530 A-19.1 Adamantylcarboxyl Adamantylcarboxyl E-531 A-19.2 Ac Ac Ac Ac E-532 A-19.2 Propionic Propionic Propionic Propionic E-533 A-19.2 Butyric Butyric Butyric Butyric E-534 A-19.2 Isobutyric Isobutyric Isobutyric Isobutyric E-535 A-19.2 Valeric Valeric Valeric Valeric E-536 A-19.2 Isovaleric Isovaleric Isovaleric Isovaleric E-537 A-20.1 Butyric Butyric Butyric E-538 A-20.1 Ac Ac Ac E-539 A-20.1 Propionic Propionic Propionic E-540 A-1 N-Phenyl H H H H CH3 chlorothionoformate E-541 A-1 Imiquimod- H H H H CH3 Succinate E-542 A-1 Resiquimod- H H H H CH3 Succinate E-543 A-1 Succinate- H H H H CH3 ethyl ester E-544 A-1 Indole-3- H H H H CH3 propionic E-545 A-1 Cyclopropanecarboxylic H H H H CH3 E-546 A-1 Cyclobutanecarboxylic H H H H CH3 E-547 A-1 Nicotinic H H H H CH3 E-548 A-1 Chenodeoxycholic H H H H CH3 E-549 A-1 Ferrocenylacetic H H H H CH3 E-550 A-1 Lipoic-S H H H H CH3 derivatives E-551 A-1 Methoxyacetic H H H H CH3 E-552 C2 (see Table 16) Butyric Butyric E-553 A-1 H Butyric Butyric H H CH3 E-554 A-1 H Ac Ac H H CH3 E-555 A-1 H Propionic Propionic H H CH3 E-556 A-1 O-Acetyl H H H CH3 Lactic E-557 A-2 (X = O) Isovaleric H H H H CH3 E-558 A-2 (X = O) Valeric H H H H CH3 E-559 A-1 Methoxyacetic Methoxyacetic Methoxyacetic Methoxyacetic H CH3 E-560 A-1 Cyclobutanecarboxylic Cyclobutanecarboxylic H H H CH3 E-561 A-1 Cyclobutanecarboxylic Cyclobutanecarboxylic Cyclobutanecarboxylic H H CH3 E-562 A-1 Nicotinic Nicotinic H H H CH3 E-563 A-1 Nicotinic Nicotinic Nicotinic H H CH3 E-564 A-2 (X = O) Ac H H H H CH3 E-566 A-10 Isobutyric Isobutyric Isobutyric H CH3 E-567 A-10 Isobutyric Isobutyric H H CH3 E-568 A-10 Isobutyric H H H CH3 E-569 A-10 Valeric Valeric Valeric H CH3 E-570 A-10 Valeric Valeric H H CH3 E-571 A-10 Valeric H H H CH3 E-572 A-10 Isovaleric Isovaleric Isovaleric H CH3 E-573 A-10 Isovaleric Isovaleric H H CH3 E-574 A-10 Isovaleric H H H CH3 E-575 A-11/E-16 Butyric H H H Butyric Butyric E-576 A-19.1 Ac H E-577 A-19.1 Propionic H E-578 A-19.1 Butyric H E-579 A-20.2 Butyric Butyric Butyric E-580 A-20.2 Ac Ac Ac E-581 A-20.2 Propionic Propionic Propionic E-582 A-12 Valeric H H H H E-583 A-12 Isovaleric H H H H E-584 A-19.3 Valeric H H E-585 A-19.3 Valeric H Ethyl E-586 A-19.3 Valeric Valeric H E-587 A-19.3 Valeric Valeric Ethyl E-588 A-19.3 Isovaleric H H E-589 A-19.3 Isovaleric H Ethyl E-590 A-19.3 Isovaleric Isovaleric H E-591 A-19.3 Isovaleric Isovaleric Ethyl E-592 A-19.3 Butyric H H E-593 A-19.3 Butyric H Ethyl E-594 A-19.3 Butyric Butyric H E-595 A-19.3 Butyric Butyric Ethyl E-596 A-17 NO.sub.2 H H H H E-597 A-17 NO.sub.2 NO.sub.2 H H H E-598 A-18 H H NO.sub.2 H H E-599 A-18 H NO.sub.2 NO.sub.2 H H E-600 A-11/E-16 H H no cladinose H H Mannose ring E-601 A-21 NO.sub.2 E-602 A-21 NO.sub.2 NO.sub.2 E-603 A-21 NO.sub.2 NO.sub.2 H NO.sub.2

[0326] Procedures

Example 1

[0327] Synthesis of E-1: Typical Nitration Procedure

##STR00035##

[0328] Method 1. Acetic acid (40 mL) and Azithromycin (5 g, 6.73 mmol) were charged in a round bottom flask. Initially, the reaction solidified which eventually during stifling, produced a homogenous solution. The resulting solution was cooled in an ice-bath. Acetic anhydride (19.8 mL, 209.5 mmol) was taken up in another reaction flask and cooled in an in ice bath. To this was added dropwise, nitric acid (2.2 mL, 46.43 mmol). After complete addition, the mixture was transferred to a dropping funnel and attached to the first reaction vessel containing the macrolide. The HNO.sub.3-Ac.sub.2O mixture was slowly added to the reaction (ca. 1 drop per second). After complete addition, the reaction was allowed to warm to room temperature where it was stirred until reaction completion (3 h). The reaction was poured onto a stirred 200 mL ice-water. Stirring was continued until the ice is completely melted. The resulting aqueous solution was neutralized at first with a saturated solution of NaHCO.sub.3, followed by pure solid NaHCO.sub.3 to pH 8 to 9. The aqueous solution was extracted with DCM (5×). The DCM extracts were dried (Na.sub.2SO.sub.4), evaporated in vacuo. The crude product was purified by column chromatography (3:1 cyclohexane, ethyl acetate, 1% triethylamine) to get compound E-1 as a white foam (30% yield).

[0329] These nitration conditions can be applied to other ALCs, and in cases where there is more than one reactive hydroxy species, selective protection is necessary.

[0330] Method 2. Hydroxyalkyl species (1 mmol) was suspended in acetonitrile in a round bottom flask while stirring (magnetic stir bar, 300 rpm). Silver nitrate (2 eq. per hydroxyl group) was added and the mixture was cooled in an ice bath. Phosgene (solution in toluene, 1 eq. per hydroxyl group) was carefully added dropwise. Immediate precipitation of silver chloride and formation of carbon dioxide indicated formation of nitro donor (caution: too quick CO.sub.2 formation may result in strong foaming. Do not seal the flask!). After a couple of minutes a yellow color was obtained and stirring was continued for 15 minutes. When ESI-MS indicated satisfying turn-over rate of starting materials the reaction was quenched by addition of methanol, converting excess nitro donor to volatile methyl nitrate. The system was diluted by addition of DCM and was subject to extraction with saturated sodium bicarbonate solution (3×). Separation of organic phase, drying over sodium sulfate and evaporation of any volatiles in vacuo yielded the product as colorless oil or beige to off-white foam.

TABLE-US-00004 TABLE 3 Nitration Examples Compound Synthesis Degree of Entry Method ALC Substitution Yield MS E-596 2 A-17 Mono nitro n.d.* 822, M + H.sup.+ E-597 2 A-17 Di nitro n.d.* 867, M + H.sup.+ 2 A-18 Oxidation n.d.* 954, M + Na.sup.+ E-598 2 A-18 Mono nitro n.d.* 999, M + Na.sup.+ E-599 2 A-18 Di nitro n.d.* 1045, M + Na.sup.+ E-601 2 A-21 Mono nitro n.d.* 793, M + H.sup.+ E-602 2 A-21 Di nitro n.d.* 838, M + H.sup.+ E-603 2 A-21 Tri nitro n.d.* 883, M + H.sup.+ n.d.* not determined

Example 2

[0331] Synthesis of E-2

##STR00036##

[0332] Compound E-1 (791 mg, 1.0 mmol) was taken up in 15 mL dichloromethane. Pyridine (89 mL, 1.1 mmol) was added and the resulting solution was cooled in an ice bath for approximately 10 minutes. At this point, a solution of acetic anhydride (113 ml, 1.2 mmol) in dichloromethane (15 mL) was added dropwise. The reaction was stirred continually at this temperature and then progressively warmed to room temperature where it was stirred overnight. The reaction was washed with a saturated solution of ammonium chloride (3×), water (3×) and dried over anhydrous Na.sub.2SO.sub.4. The solvent was evaporated in vacuo. Co-evaporation with toluene is necessary to remove residual pyridine from the system. This was followed by re-dissolving the residue in DCM and solvent evaporation twice to produce a white foam, which was dried under high-vacuum to produce E-2 (631 mg, 76%).

Example 3

[0333] Synthesis of E-3

##STR00037##

[0334] E-3 was synthesized using standard nitration procedure described above starting from E-20.

Example 4

[0335] Synthesis of E-4

##STR00038##

[0336] A solution of E-3 (1 mmol) and pyridine (1 mmol) in DCM (10 ml) was treated with acetic anhydride (1.5 mmol) at ambient temperature. Stirring was continued until TLC (acetone-cyclohexane 1:3) indicated complete consumption of the starting materials. The system was extracted with an aqueous solution of NH4Cl (2× 10 ml) and water (3× 10 ml). After drying over Na.sub.2SO.sub.4 all volatile components were evaporated in vacuo, traces of pyridine species were removed by co-evaporation with toluene (2×). The product E-4 was a colorless oil (41%).

Example 5

[0337] Synthesis of E-5

##STR00039##

[0338] Method 1. A solution of propionic acid (1.2 mmol) in 1.2-dichloroethane was treated with 1-Ethyl-3-(3-dimethyl-aminopropyl)carbodiimid (1.2 mmol) in the presence of a catalytical amount of DMAP for 30 min. at ambient temperature. Temperature was raised to 55°C., E-1 (1 mmol) was added and stirring was continued until TLC (acetone-cyclohexane 1:3) indicated complete consumption of the starting materials. The system was extracted with water (3× 10 ml). After drying over Na.sub.2SO.sub.4 all volatile components were evaporated in vacuo, traces of pyridine species were removed by co-evaporation with toluene (2×). The crude products were purified by column chromatography (acetone-cyclohexane 1:3), yielding the product E-5 as white amorphous foam (39%).

[0339] Alternative Synthesis: Compound E-1 (310 mg, 0.39 mmol) was taken up in dichloromethane (15 mL). At which point, pyridine (32 mL, 0.39 mmol) was added. The solution was stirred for 5 minutes, at which time, propionyl anhydride (51 mL, 0.40 mmol) was added. The reaction was allowed to stir at room temperature for 72 h. An additional propionyl anhydride (0.1 eq) was added and the reaction monitored until complete disappearance of starting material was observed (MS, reaction may take up to 1 week). The reaction was washed successively with a saturated aqueous solution of NH.sub.4Cl (3×) and H.sub.2O (3×). The organic phase was dried over anhydrous Na.sub.2SO.sub.4 and evaporated in vacuo. Co-evaporation with toluene is necessary to remove residual pyridine from the system. This was followed by re-dissolving the residue in DCM and solvent evaporation twice to produce E-5 as a white foam (210 mg, 64%).

[0340] Method 3: A-1 (300 mg, 0.40 mmol), was dissolved in DCM (10 mL); to this solution was added TEA (279 μL, 5 eq) and propionylchloride (175 μL, 5 eq) subsequently and the mixture was stirred overnight at room temperature. Additional TEA (112 μL, 2 eq) and propionyl chloride (70 μL, 2 eq) were added and again mixture was stirred overnight at room temperature. Once more additional TEA (112 μL, 2 eq) and propionyl chloride (70 μL, 2 eq) were added and stirring at room temperature was continued overnight. TEA (344 μL, 9 eq) and propionyl chloride (314 μL, 9 eq) were added and the mixture was stirred at room temperature for 5 days. The reaction mixture was washed with aqueous Na.sub.2CO.sub.3-solution (3×, 10%) and water (3×), dried, concentrated to dryness, and dried at the oil pump. ESI-MS (positive) showed tri- and tetra-propionylation.

[0341] Method 4

[0342] Propionic acid (4 eq) was taken up in 5 mL dichloromethane (DCM). Compound A-16 (0.5 mmol) and 4-dimethylaminopyridine (DMAP) (4.4 eq) were added and the resulting solution was cooled in an ice bath for approximately 10 minutes. At this point, dicyclohexylcarbodiimide (DCC) (4.4 eq) was added slowly. The reaction was stirred continually at this temperature for 5 minutes and then progressively warmed to room temperature where it was stirred overnight. Dicyclohexylurea (DCC) that was formed during the reaction is filtered off and discarded. The filtrate was collected and then washed with a saturated solution of sodium hydrogencarbonate (3×), water (1×) and dried over anhydrous Na.sub.2SO.sub.4. The solvent was evaporated in vacuo. This was followed by re-dissolving the residue in a small volume of methanol. The solution was transported dropwise into ice-cold water (2× volume of methanol) and stored in the freezer overnight. The precipitated product was filtered off and dried under high-vacuum to produce

TABLE-US-00005 TABLE 4 Propionylation Examples Reaction MS Compound Synthesis Substituent Condition Degree of m/z Entry Method ALC equivalent (i.e. Workup) Substitution Yield ([M + H].sup.+) E-5 1 A-1 1.1 as described 1 64% 850.3 above  E-18 See A-1 N/A 1 80% 805.5 example 21  E-419 4  A-16 4 1 50% 791.3  E-420 2 (based on 847.1  E-421 3 tri-ester 903.0  E-52 3 A-1 18 3 Not 917.9  E-51 4 determined 973.8

Example 6

[0343] Synthesis of E-8

##STR00040##

[0344] Erythromycin oxime was nitrated as described in the general procedure, followed by subsequent acetylation of crude nitration product with acetic anhydride in DCM with pyridine as catalyst. Column chromatography (acetone-cyclohexane 1:3) furnished the product E-8 as white amorphous foam (39%, two steps).

Example 7

[0345] Synthesis of E-10

##STR00041##

[0346] A solution of E-9 (1 mmol) in MeOH (20 ml) was vigorously stirred at 50° C. until TLC (acetone-cyclohexane 1:3) indicated complete deacetylation of the starting materials. Volatile components were removed in vacuo, the product E-10 was obtained as white amorphous foam (81%).

Example 8

[0347] Synthesis of E-11

##STR00042##

[0348] Azithromycin was nitrated as described above. After desired mono-nitration MeOH was added to the reaction mixture and stirring was continued for one further hour. Thus in situ generated methyl nitrate acted as methylating agent transferring one methyl group to 11-O-position of the macrolide at ambient temperature. Standard aqueous workup with subsequent purification by column chromatography (acetone-cyclohexane 1:3) delivered methylated macrolide nitrate E-11 as white amorphous solid (63%, two steps).

Example 9

[0349] Synthesis of E-16

##STR00043##

[0350] Azithromycin (20.0 g; 26.7 mmol) was dissolved in 120 ml of MeOH. NaHCO.sub.3 (6.0 g; 71.5 mmol) was added, followed by a solution of K.sub.2CO.sub.3 (12.0 g; in water (80 ml; cooled down to RT), and finally iodine (6.3 g; 24.8 mmol). The mixture was stirred vigorously at ambient temperature until the dark color had disappeared. A second batch of iodine (6.3 g; 24.8 mmol) and K.sub.2CO.sub.3 carbonate (4.2 g; 30 mmol) were added. The procedure [addition of iodine 6.3 g and K.sub.2CO.sub.3 (4.2 g; 30 mmol)] was repeated until MS showed (almost) full conversion. Sodium bisulfite was added to remove excess oxidants, and all volatiles were evaporated. The solid residue was finely ground and extensively extracted via Soxhlet extraction with acetonitrile. The extract was concentrated to ca. 75 ml and left standing at ambient temperature at least for 1 day and subsequently for another day in the fridge. All solids were collected and recrystallized from MeOH spiked with ca. 1 to 2 ml of water. Crystallization proceeded for about 3 days in an open vessel to yield 10 g (51%) of 3′-N-demethyl-azithromycin (E-16). A second crop can be obtained from the mother liquors.

Example 10

[0351] Synthesis of E-20

##STR00044##

[0352] A solution of E-16 (5 mmol) in DMSO (20 ml) was treated with decyl bromide (6 mmol) at ambient temperature. Stirring continued for 12 h until TLC (acetone-cyclohexane 1:3, 1% Et.sub.3N) indicated consumption of starting materials. The system was diluted with EtOAc. (50 ml) and extracted with water (3× 30 ml). The organic phase was dried over sodium sulfate. Evaporation of the solvent and drying at vacuum yielded E-20 as white amorphous foam (61%).

Example 11

[0353] Synthesis of E-25

##STR00045##

[0354] A solution of azithromycin (13 mmol) and pyridine (13 mmol) in DCM (80 ml) was cooled to 0° C. in an ice bath. A solution of acetic anhydride (14 mmol) in DCM (20 ml) was slowly added to the system. Afterwards the reaction mixture was allowed to warm up to ambient temperature and stirring was continued until TLC (acetone-cyclohexane 1:3, 1% Et.sub.3N) indicated complete consumption of the starting materials. The system was extracted with an aqueous solution of NH.sub.4Cl (2× 50 ml) and water (3× 50 ml). After drying over Na.sub.2SO.sub.4 all volatile components were evaporated in vacuo, traces of pyridine were removed by coevaporation with toluene (2×). The product E-25 was a white amorphous solid (67%).

Example 12

[0355] Synthesis of E-22

##STR00046##

[0356] A solution of E-25 (4 mmol) in dry THF (30 ml) was cooled to 0° C. in an ice bath. A solution of propargyl bromide (4.4 mmol, 80% in toluene) was added slowly to the system. Afterwards the reaction mixture was allowed to warm up to ambient temperature and stirring was continued until TLC (acetone-cyclohexane 1:3, 1% Et.sub.3N) indicated complete consumption of the starting materials. The system was diluted with EtOAc (50 ml) and extracted with water (3× 50 ml). After drying over Na.sub.2SO.sub.4 volatile components were evaporated in vacuo. Purification by column chromatography (acetone-cyclohexane 1:3, 1% Et.sub.3N) furnished E-22 as white amorphous powder (57%).

Example 13

[0357] Synthesis of E-12

##STR00047##

[0358] A solution of E-22 (0.5 mmol), I-5 (0.5 mmol) and DIPEA (1 mmol) in toluene (5 ml) was treated with triethylphosphito copper(I) iodide complex (0.05 mmol) at ambient temperature. Stirring was continued until TLC (ethyl acetate-cyclohexane 1:1) indicated complete consumption of the starting materials. The mixture was concentrated in vacuo and purified by column chromatography (acetone-cyclohexane 1:1.fwdarw.acetone). The product E-12 was obtained as colorless foam (32%).

Example 14

[0359] Synthesis of E-23

##STR00048##

[0360] A solution of E-25 (5 mmol) and pyridine (5 mmol) in DOA (40 ml) was treated with acetic anhydride (7 mmol) at ambient temperature. Stirring was continued until TLC (acetone cyclohexane 1:3, 1% Et.sub.3N) indicated complete consumption of the starting materials. The system was extracted with an aqueous solution of NH.sub.4Cl (2× 20 ml) and water (3× 30 ml). After drying over Na.sub.2SO.sub.4 all volatile components were evaporated in vacuo, traces of pyridine species were removed by co-evaporation with toluene (2×). The product E-23 was a white amorphous powder (54%).

Example 15

[0361] Synthesis of E-17

##STR00049##

[0362] A solution of E-23 (2 mmol) in MeOH (30 ml) was vigorously stirred at 50° C. until TLC (acetone-cyclohexane 1:3, 1% Et.sub.3N) indicated complete deacetylation of the starting materials. Volatile components were removed in vacuo, the product E-17 was obtained as white amorphous foam (73%).

[0363] Alternative Synthesis: Compound E-2 (619 mg, 0.74 mmol) was charged in a round bottom flask. To this was added a solution of acetic acid/methanol (2:1, 15 mL). To the stirred solution, was added Zn powder (368 mg, 5.63 mmol, 7.6 eq). The resulting suspension was stirred at room temperature and progressively monitoring the disappearance of the starting material (approx. 3 h). The suspension was filtered and the filtrate evaporated in vacuo. The residue was taken up in dichloromethane (15 mL) producing some white precipitate. The precipitate was filtered off. The dichloromethane filtrate was washed with 10% aqueous Na.sub.2CO.sub.3 solution (2×), water (1×) and dried over anhydrous Na.sub.2SO.sub.4. The solvent was evaporated in vacuo producing E-17 as a white solid (475 mg, 81% yield).

Example 16

[0364] Synthesis of E-26

##STR00050##

[0365] A solution of azithromycin (13 mmol) and pyridine (13 mmol) in DCM (80 ml) was cooled to 0° C. in an ice bath. A solution of benzoyl chloride (14 mmol) in DCM (20 ml) was slowly added to the system. Afterwards the reaction mixture was allowed to warm up to ambient temperature and stirring was continued until TLC (acetone-cyclohexane 1:3, 1% Et.sub.3N) indicated complete consumption of the starting materials. The system was extracted with an aqueous solution of NH.sub.4Cl (2× 50 ml) and water (3× 50 ml). After drying over Na.sub.2SO.sub.4 all volatile components were evaporated in vacuo, traces of pyridine species were removed by co-evaporation with toluene (2×). Column chromatography (acetone-cyclohexane 1:3, 1% Et.sub.3N) yielded the product E-26 as a white amorphous foam (44%).

Example 17

[0366] Synthesis of E-13

##STR00051##

[0367] A solution of E-5 (1 mmol) and propionic acid (1.2 mmol) in 1,2-dichloroethane (3 ml) was treated portion wise with EDCI (3× 0.8 mmol) and DMAP (1 mmol). The mixture was heated to 55° C. and stirred for 6 days. After TLC (acetone-cyclohexane 1:3) indicated complete consumption of the starting materials the mixture was diluted with EtOAc (30 ml) and extracted with water (3× 20 ml). After drying over Na.sub.2SO.sub.4 the organic phase was evaporated and the remains were purified by column chromatography (acetone-cyclohexane 1:3). The product E-13 was obtained as colorless amorphous foam (42%).

Example 18

[0368] Synthesis of E-14

##STR00052##

[0369] 3′-N-desmethyl-azithromycin (I-1) (300 mg; 0.41 mmol) and 1-bromo-3-nitrooxy-propane.sup.[17] (90 mg, 0.49 mmol) were dissolved in dry DMSO (1.2 ml). The mixture was shaken at 23° C. for 3 hours. Afterwards additional 1-bromo-3-nitrooxy-propane (90 mg, 0.49 mmol) was added and shaking was continued for another hour. Once again, 1-bromo-3-nitrooxy-propane (90 mg, 0.49 mmol) was added, the reaction mixture was shaken for additional 90 min., and then kept in the freezer (−16° C.) over night. The next morning additional 1-bromo-3-nitrooxy-propane (90 mg, 0.49 mmol) was added and the mixture was shaken for one hour. Water and DCM were added; after extraction, the organic phase was dried (Na.sub.2SO.sub.4) and concentrated to dryness (without heating). The crude product was purified by column chromatography (eluent: CHCl.sub.3:Isopropanol: NH.sub.3 (7 M in MeOH 30:1:1).

Example 19

[0370] Synthesis of (I-5)

##STR00053##

[0371] Azido-β-D-Glucopyranoside is synthesized from corresponding sugar acetate as is known to literature.sup.[16]. After removal of any protective groups sugar azide is nitrated following above procedure. Due to its high explosive risk the substance is always kept as DCM solution and is stashed in the refrigerator.

Example 20

[0372] Synthesis of E-24

##STR00054##

[0373] Compound E-1 (405 mg, 0.51 mmol) was taken up in dichloromethane (15 mL). The resulting solution was cooled to 0° C. After 5 minutes, butyryl chloride (60 mL, 61.8 mg, 0.58 mmol, 1.1 eq.) was added. The reaction was stirred for 10 min. at this temperature, at which point, the reaction was allowed to warm to room temperature, where it was stirred until reaction completion (2 h, or upon continuous monitoring). The reaction was washed with a 10% Na.sub.2CO.sub.3 aq. solution (3×) followed by H.sub.2O (3×), dried over anhydrous Na.sub.2SO.sub.4 and evaporated in vacuo to give E-24 as a white foam (332 mg, 75% yield).

Example 21

[0374] Synthesis of E-18

##STR00055##

[0375] Compound E-5 (CSY 1076) (126 mg, 0.15 mmol) was charged in a round bottom flask. To this was added a solution of acetic acid/methanol (2:1, 12 mL). To the stirred solution, was added Zn powder (74 mg, 1.12 mmol, 7.6 eq). The resulting suspension was stiffed at room temperature and progressively monitoring the disappearance of the starting material (approx. 3 h). The suspension was filtered and the filtrate evaporated in vacuo. The residue was taken up in dichloromethane (10 mL) producing some white precipitate. The precipitate was filtered off. The dichloromethane filtrate was washed with 10% aqueous Na.sub.2CO.sub.3 solution (2×), water (1×) and dried over anhydrous Na.sub.2SO.sub.4. The solvent was evaporated in vacuo producing E-18 as a white solid (96 mg, 80% yield).

[0376] The reduction conditions for removal of the NO.sub.2 group was applied to the syntheses of E-19 from E-24 providing the desired product in 78% yield.

Example 22

[0377] Synthesis of E-19

##STR00056##

[0378] Compound E-24 CSY 4636 (100 mg, 0.12 mmol) was charged in a round bottom flask. To this was added a solution of acetic acid/methanol (2:1, 12 mL). To the stirred solution, was added Zn powder (88 mg, 1.35 mmol, 11.6 eq). The resulting suspension was stirred at room temperature and progressively monitoring the disappearance of the starting material (approx. 3 h). The suspension was filtered and the filtrate evaporated in vacuo. The residue was taken up in dichloromethane (15 mL) producing some white precipitate. The precipitate was filtered off. The dichloromethane filtrate was washed with 10% aqueous Na.sub.2CO.sub.3 solution (2×), water (1×) and dried over anhydrous Na.sub.2SO.sub.4. The solvent was evaporated in vacuo producing E-19 as transparent gel (74 mg, 78% yield).

Example 23

[0379] Synthesis of E-81

##STR00057##

[0380] 250 mg of 2′-(2-Mercaptoethoxy)carbonyl-3-decladinosylazithromycin and 250 mg of ammonium polysulfide are mixed with 10 ml of degassed and argonized glacial acetic acid and stirred with exclusion of oxygen for 12 h. All volatiles are removed in vacuo, and the residue is extracted with oxygen free saturated aqueous sodium hydrogen carbonate solution 3 times. The residue is washed with water (oxygen free), dried in vacuo and used as such.

Example 24

[0381] Synthesis of E-82

##STR00058##

[0382] 250 mg of 2′-(2-Mercaptoethoxy)carbonyl-3-decladinosylazithromycin are dissolved in a mixture of 5 ml of tert. butanol and 5 ml of dichloromethane. 250 μl of tert. butylnitrite are added, and the mixture is stirred for 48 h with exclusion of light. All volatiles are removed i.v. keeping the temperature below 20° C. and light excluded. The red residue is used as such

Example 25

[0383] Synthesis of E-74

##STR00059##

[0384] 380 mg of Azithromycin are dissolved with 20 ml of DMF and cooled in an ice bath. 41 ml of pyridine and then 85 mg (1.1 eq.) of rac-2-acetoxy propionyl chloride, dissolved with 1 ml of dichloromethane, are added and the mixture is allowed to warm up to room temperature with stirring. When mass spectrometry indicates consumption of the macrolide, the mixture is diluted with 50 ml of ethyl acetate, extracted twice with water, 3 times with saturated aqueous sodium hydrogen carbonate solution, once again with water and brine, each, and dried over sodium sulfate. After evaporation and vacuum drying, the diastereomeric mixture (1:1) of target compound E-74 remains as a slightly yellowish foam.

[0385] Yield: 385 mg

[0386] MS: m/z=863.5 ([M+H].sup.−)

[0387] The same procedure can be applied in preparing E-77, E-83, E-84, E-85 and E-86.

TABLE-US-00006 TABLE 5 Typical Products from the Acylation Procedures Entry Acylating agent Product structure Yield [%] ([M + H].sup.+ E-83 O-Phenyl chlorothionoformate [00060] 66 885.7 E-85 2- Bromoethylchloroformate [00061] 82 899.6 E-84 Hexanoylchloride [00062] 93 847.6 E-77 Lipoyl chloride [00063] 72 936.8 E-86 Allylchloroformate [00064] 82 833.5 E-540 O-Phenyl chlorothionoformate [00065] 23 884 E-26 Benzoyl [00066] 44 853

Example 26

[0388] General Procedure: Synthesis of n-Butyryl-Propanolol, Compound E-133

##STR00067##

[0389] Propranolol HCl (200 mg, 0.68 mmol) was taken up in dichloromethane (4 mL). To this was added dropwise butyryl chloride (69 μL, 0.71 mmol) and the reaction was stirred at room temperature for 1 hour. To the reaction was added triethylamine (194 μL, 1.4 mmol). After 30 min, additional butyryl chloride (30 μL, 0.3 mmol) was added. Reaction was monitored by the disappearance of starting material. The reaction was stopped after 20 min by the addition of 10 mL 10% aqueous Na.sub.2CO.sub.3 solution. The two phases were stirred for 5 min separated. The organic layer was washed successively with 10% aqueous Na.sub.2CO.sub.3 (1×), H.sub.2O (1×) and saturated aq. NaCl (1×), dried with Na2SO4 and evaporated in vacuo to get an oil film. 5 mL HCl in Et.sub.2O (2M) and 1 mL MeOH was added and stirred for 5 min, then evaporated in vacuo and dried with the vacuum pump under nitrogen for 1 hour to get a brown oil (yield 91%).

Example 27

[0390] General Procedure: Synthesis of n-Butyryl-Hydroxychloroquine, Compound E-89

##STR00068##

[0391] Hydroxychloroquine sulfate (1099 mg, 2.53 mmol) was charged into a round bottom flask. H.sub.2O (10 mL) and dichloromethane (10 mL) were added. Pyridine (412 μL, 5.1 mmol) was added and the reaction stirred vigorously for 5 min. Butyric anhydride (420 μL, 2.65 mmol) was added and the reaction stirred at room temperature for 3 h. The phases were separated and the dichloromethane layer was washed successively with a saturated aqueous NH.sub.4Cl solution (2×15 mL), H.sub.2O (2×10 mL), dried over Na.sub.2SO.sub.4 and evaporated in vacuo. Co-evaporation with toluene is necessary to remove residual pyridine from the system. This was followed by re-dissolving the residue in DCM and solvent evaporation twice to produce a yellow oil (93 mg, 9% yield).

[0392] This procedure can be applied for the synthesis of the following compounds E-87 to E-88 E-90 to E-97.

Example 28

[0393] Typical methylation reaction of Hydroxychloroquine or Propranolol was achieved using Eschweiler-Clarke-methylation reaction. Acylation reactions were carried out using typical procedures described above.

Example 29

[0394] Compounds, prepared by reacting a carrier molecule with acylating agents.

[0395] These carrier molecules are prepared by reacting symmetric or unsymmetric di- or poly-epoxides with secondary amines, thus containing the common structural element of 2 or more alcohols, vicinally neighbored by a tertiary amine.

[0396] Alternatively, carrier molecules can be prepared by reacting epoxides with diethanolamine. The reaction products are containing 2-hydroxy tertiary amines.

TABLE-US-00007 TABLE 6 Examples for di- or polyepoxides No. of epoxide Entry Name CAS-No. functions A 1,3-Butadiene diepoxide 1464-53-5 2 B 1,4-Butandiole diglycidylether 2425-79-8 2 C N,N-Diglycidylaniline 2095-06-9 2 D Resorcinol diglycidylether 101-90-6 2 E Ethylen glycol diglycidylether 2224-15-9 2 F 1,7-octadienediepoxide 2426-07-5 2 G Diglycidyl ether 2238-07-5 2 H 1,2,4,5,9,10-Triepoxydecane 52338-90-6 3 1 N,N-Diglycidyl-4-glycidyloxyaniline 5026-74-4 3 J Poly(ethylene glycol) diglycidyl ether 72207-80-8 2 (average M.sub.n 500) K Glycerol diglycidylether 72207-80-8 2 L 4,4′-Methylenebis(N,N- 28768-32-3 4 diglycidylaniline) M Bis[4-(glycidyloxy)phenyl]methane 2095-03-6 2

TABLE-US-00008 TABLE 7 Examples for secondary amines Entry Name CAS-No. 1 Dimethylamine 2 Morpholine 3 4-Methylpiperazine 4 Piperidine 5 1,2,3,4-Tetrahydroisoquinoline 91-21-4 6 Diethylamine 7 Dioctylamine 8 Diethanolamine 9 Sarcosine methyl ester hydrochloride 13515-93-0 10 (R)-pyrrolidine-2-carboxylic acid methyl ester (Prolin methylester) 11 Ethyl 1,4-diazepan-1-ylacetate dihydrochloride

TABLE-US-00009 TABLE 8 Structural examples for carrier molecules Combination of poly-epoxide and amine Structural Example H1 [00069] G5 [00070] C8 [00071] A-7-C [00072] B2 [00073]

[0397] The 2-aminoalcohols of these carriers can be esterified to short chain carboxylic acids or nitric acid. One molecule can contain esters of different of these acids. Examples for short chain carboxylic acids are:

[0398] Acetic acid, propionic acid, butyric acid, isobutyric acid, valeric acid, 2-methylbutyric acid, 3-methylbutryric acid, lactic acid, pyruvic acid, 3-phenylpropionic acid, succinic acid, maleic acid, fumaric acid, malic acid, lactic acid butyrate (lactic acid butanioate), 2-acetoxy propionic acid, mandelic acid, benzoic acid.

[0399] Structural Examples of such Esters are:

##STR00074##

[0400] Alternatively, these carrier molecules are prepared by reacting symmetric or unsymmetric di- or polyamines with epoxides, thus containing the common structural element of 2 or more alcohols, vicinally neighboured by a tertiary amine.

TABLE-US-00010 TABLE 9 Examples for di- or polyamines No. of reactive Entry Name CAS. No. amines N Spermine 71-44-3 4 O Spermidine 124-20-9 3 P Piperazine 110-85-0 2 Q 1-(2-Aminoethyl)piperazine 140-31-8 2 R L-Lysine 56-87-1 2 S Homopiperazine 505-66-8 2 T 1,3-Diamino-2-propanol 616-29-5 2 U 1,3,5-Triamino-1,3,5-trideoxy-cis-inositol 6988-69-8 3 trihydrochloride V 1,2,3,4-Tetrahydroquinoxaline 3476-89-9 2 W Tetraethylenepentamine 112-57-2 5

TABLE-US-00011 TABLE 10 Examples for epoxides Entry Name CAS No. 12 Propylene oxide 75-56-9 13 Cyclohexen oxide 286-20-4 14 Ethyl 2,3-epoxypropionate 4660-80-4 15 Styrene oxide 96-09-3 16 Glycidol 556-52-5

TABLE-US-00012 TABLE 11 Structural examples for carrier molecules Combination of poly-epoxide and amine Structural Example Q12 [00075] Q13 [00076] R16 [00077] S14 [00078] T15 [00079]

[0401] The 2-aminoalcohols of these carriers can be esterified to short chain carboxylic acids or nitric acid. One molecule can contain esters of different of these acids. Examples for short chain carboxylic acids are:

[0402] Acetic acid, propionic acid, butyric acid, isobutyric acid, valeric acid, 2-methylbutyric acid, 3-methylbutryric acid, lactic acid, pyruvic acid, 3-phenylpropionic acid, succinic acid, maleic acid, fumaric acid, malic acid, lactic acid butyrate (lactic acid butanoate), 2-acetoxy propionic acid, mandelic acid, benzoic acid.

[0403] Structural Examples of such Esters are:

##STR00080##

Example 30

[0404] Synthesis of Rac. 2′-Deoxy-2′-S-Thioacetyl ppropranolol.sup.[18]

##STR00081##

[0405] Diethyl azodicarboxylate (DEAD, 10 mmol) is added to a stirred (magnetic stirrer, 300 rpm) solution of triphenylphosphin (10 mmol) in dry THF (25 mL) at 0° C. and treatment is continued for 30 min. Propranolol (5 mmol) and thioacetic acid (10 mmol) both dissolved in THF (10 mL) are added dropwise and stirring is continued for 1 h at 0° C. and further 2 h at ambient temperature. Any precipitates are filtered off, the remaining solution is concentrated in vacuo and desired product is isolated by column chromatography (silica gel, cyclohexane-ethyl acetate).

[0406] Synthesis or Rac. 2′-Deoxy-2′-S-Thio Propranolol Sodium Salt

##STR00082##

[0407] rac. 2′-Deoxy-2′-S-thioacetyl propranolol (10 mmol) is dissolved in methanol (20 mL) while stirring (magnetic stirrer, 300 rpm) and sodium methoxide (10 mmol) is added at 0° C. The system is allowed to warm up to ambient temperature and treatment is continued until TLC (cyclohexane-ethyl acetate) indicates full conversion of starting materials. Afterwards the reaction mixture is rinsed into ice cold diethyl ether (100 mL). Any precipitates are filtered off and are dried in vacuo to yield a white to slightly yellow product.

Example 31

[0408] Synthesis of Rac. 2-O-Nitrolactic acid

##STR00083##

[0409] Lactic acid (10 mmol) is suspended in acetonitrile (20 mL) in a 3-necked round bottom flask and is cooled to 0° C. in an ice bath while stirring (300 rpm). Diphosgene is added (5 mmol) followed by careful dropwise addition of silver nitrate solution (20 mmol, dissolved in acetonitrile). The mixture is stirred for 30 min at 0° C., subsequently is allowed to warm up to ambient temperature and stirring is continued for further 30 min. Afterwards any precipitates are filtered off and the mixture is carefully concentrated in vacuo, As crude products are likely to be explosive compounds the system was not fully dried but taken up in THF (10 mL) to be immediately used in the following step.

[0410] Synthesis of Rac. 2′-O-(2-O-Nitrolactyl) Propranolol—E-42

##STR00084##

[0411] Diethyl azodicarboxylat (DEAD, 10 mmol) is added to a stirred (magnetic stirrer, 300 rpm) solution of triphenylphosphin (10 mmol) in dry THF (25 mL) at 0° C. and treatment is continued for 30 min. Propranolol (5 mmol) and rac. 2-O-nitrolactic acid (10 mmol) both dissolved in THF (10 mL) are added dropwise and stirring is continued for 1 h at 0° C. and further 2 hours at ambient temperature. Any precipitates are filtered off, the remaining solution is concentrated in vacuo and desired product is isolated by column chromatography (silica gel, cyclohexane-ethyl acetate).

Example 32

[0412] Rac. Bis-(2′-Deoxy-2′-S-S-Disulfido Propranolol) (Putative).sup.[19]

##STR00085##

[0413] A round bottom flask is charged with ethyl acetate (10 mL), graphite (3 g), iodine (0.5 mmol) and cerium(III) chloride heptahydrate (1 mmol). The mixture is stirred for 10 min at ambient temperature, followed by addition of rac. 2′-deoxy-2′-S-thio propranolol sodium salt (10 mmol). Treatment is continued until TLC (cyclohexane-ethyl acetate) indicates full conversion of starting materials. After completion the system is further diluted with additional ethyl acetate (250 mL) and is washed with a saturated solution of aqueous sodium thiosulfate, water, and is dried over sodium sulfate. Upon filtration any volatiles are removed in vacuo and the residue is subject to column chromatography (silica gel, cyclohexane-ethyl acetate).

[0414] Mixed disulfides may also be accessible in this way but require slight alterations.

Example 33

[0415] Typical Example or Synthesizing ALC Cores A-8 and A-9

[0416] Synthesis of 2-(4-Pyridyl)-3-Amino-4-(3-Methoxyphenyl)Carbonyl Pyrazole

##STR00086##

[0417] 10.96 g of 4-(N-phenyl)amino-3-(3-methoxyphenyl)carbonylacrylonitrile and 6.27 g of 4-pyridylhydrazine hydrochloride are combined with 6.2 ml of triethylamine in 140 ml of ethanol, flushed with argon and heated to reflux for 5 h. The mixture is concentrated to 50 ml and diluted with 200 ml of cyclohexane. The precipitate is filtered off and washed with diethyl ether, until no further colour is extracted any more. The remaining solid is dissolved with a mixture of dichloromethane and water (300 ml each). The organic phase is dried with brine and sodium sulfate and concentrated to dryness.

[0418] Yield: 7.07 g (61%); MS: m/z=295 ([M+H]+)

[0419] Synthesis of 2-(4-Pyridyl)-3-Amino-4-(3-Hydroxyphenyl)Carbonyl Pyrazole

##STR00087##

[0420] 2.6 g of 2-(4-Pyridyl)-3-amino-4-(3-methoxyphenyl)carbonylpyrazole are suspended in 10 ml of a solution of 33% hydrobromic acid in acetic acid. The mixture is heated to 70° C. for 16 h. After cooling, the reaction mix is poured into 150 ml of water. The precipitate is filtered off, washed with saturated aqueous sodium hydrogen carbonate solution (twice) and water, and dissolved in 10 ml of a solution of ammonia in methanol (7M). After 30 min, all volatiles are removed by evaporation, and the residue is dissolved in 50 ml of boiling methanol. The product is precipitated by pouring into 250 ml of water. Filtration and drying yield 2.35 g of an off white powder.

[0421] Synthesis of 2-(4-Pyridyl)-3-Amino-4-(3-[2,3-Dihydroxypropyloxy]Phenyl)-Carbonyl Pyrazole

##STR00088##

[0422] 2.09 g of 2-(4-Pyridyl)-3-amino-4-(3-hydroxyphenyl)carbonyl pyrazole are dissolved with 25 ml of dimethylformamide. 3.5 g of potassium carbonate and 580 mg of glycidol are added. The mixture is kept stirring at 60° C. for 18 h. The reaction mixture is partitioned between water and ethyl acetate. The organic phase is washed with water, dried with brine and sodium sulfate, and concentrated i.v. The residue is subjected to preparative HPLC to yield 1.4 g of the product.

[0423] Synthesis of 2-(4-Pyridyl)-3-Amino-4-(3-[2,3-Di{Butyroyloxy}Propyloxy]Phenyl)Carbonyl Pyrazole E-199

##STR00089##

[0424] 500 mg of 2-(4-pyridyl)-3-amino-4-(3-[2,3-dihydroxypropyloxy]phenyl)carbonylpyrazole are dissolved with 5 ml of pyridine. 500 μl of butyric acid anhydride are added, and the mixture is stirred at 50° C. over night. 1 ml of methanol is added, and the mixture is stirred for further 30 min. The reaction is allowed to reach room temperature and partitioned between water and ethyl acetate. The organic phase is extracted with water 5 times, then with brine and dried over sodium sulfate. After evaporation of all volatiles, the product is purified by chromatography over silica gel.

[0425] Yield: 290 mg

[0426] The same conditions can be applied to synthesis of 2-(4-fluorophenyl)-3-amino-4-(3-[2,3-di{butyroyloxy}propyloxy]phenyl)carbonylpyrazole E-210

##STR00090##

Example 34

[0427] Synthesis of Chenodeoxycholic Acid Azithromycin-2′-Ester

##STR00091##

[0428] 1 g of Chenodeoxy cholic acid is dissolved with 50 ml of dry dichloromethane and cooled in an ice bath. 500 mg of carbonyl diimidazole are added, and the mixture is stirred for 2 h, while reaching room temperature. 2 g of Azithromycin are added, and the mixture is stirred for 72 h. The mixture is extracted with water (3×) and then with 5% citric acid. The citric acid phase is washed with dichloromethane (2×). It is then vigorously stirred with ethyl acetate, while portions of sodium hydrogencarbonate are added, so that gas evolution is under control. When no gas is developed any more, the organic phase is isolated, washed with brine and dried over sodium sulfate. Concentration and chromatography with a gradient starting at 10% of acetone in cyclohexane (always containing 0.2% of triethylamine) yields 350 mg of the desired product.

Example 35

[0429] Synthesis of 2′-(Succinyl-1-Hydroxymethylferrocene)-11-Nitro-Azithromycin

##STR00092##

[0430] Compound E-10 (200 mg, 0.25 mmol) was dissolved in dry dichloromethane (5 mL). To this was added subsequently, 4-dimethylaminopyridine (4-DMAP, 3 mg, 0.25 mmol. 0.1 eq.) and succinic anhydride (28 mg, 0.28 mmol). The reaction was stirred overnight at room temperature. The solvent was removed in vacuo and the resulting white amorphous foam was used directly for the next step.

[0431] Fresh dry dichloromethane (5 mL) was added to the resulting foam, followed by 1-Hydroxy-methylferrocene (60 mg, 0.28 mmol, 1.1 eq.). The reaction was cooled to 0° C. and to this was added EDCI (96 mg, 0.5 mmol, 2 eq.). The reaction was allowed to progressively warm to room temperature where it was stirred overnight. Additional dichloromethane (20 mL) was added and washed several times with saturated aqueous ammonium chloride, brine (2×), dried under anhydrous Na.sub.2SO.sub.4 and the solvent removed in vacuo. The resulting crude product was purified by chromatography with a gradient starting at 10% of acetone in cyclohexane (0.2% Et.sub.3N) yields 150 mg of the desired product (53%).

[0432] Similarly, the following compound may be obtained using the procedure above starting from compound E-19

##STR00093##

Example 36

[0433] Activity of substances in the inhibition of growth of bacteria. Bacteria including the species Escherichia coli, Bacillus pumilus, Salmonella sp., Micrococcus luteus and Staphylococcus carnosus are cultured in appropriate media (Luria broth for all except S. canosus). Overnight cultures are mixed with fresh medium to reach an optical density at 600 nM of ca. 0.1 AU. These cultures are mixed with solutions of substances to be tested at concentrations ranging from 100 μM to 0.05 μM in a microtitre plate. The growth of the culture is monitored by measuring the optical density at various times after the addition of the inhibitor. Reduction in the rate of increase in optical density corresponds to an inhibition of bacterial growth. In the following tables, the activity of various of the test substances may be observed by reductions in optical density relative to untreated control cultures. The data are summarized in Table 3 and Table 4.

TABLE-US-00013 TABLE 12 Inhibition of growth of Staphylococcus carnosus by compounds after 9-20 h: Conc. Absorbance of culture medium at 600 nm (μM): 100 50 25 13 6 3 2 0.8 E-2 0.395 0.425 0.456 0.542 0.626 0815 0.766 0.760 E-5 0.302 0.347 0.403 0.452 0.525 0.805 0.726 0.819  E-13 0.421 0.437 0.282 0.488 0.530 0.668 0.755 0.765 E-1 0.157 0.096 0.078 0.068 0.625 0.864 0.856 0.902  E-12 0.494 0.523 0.574 0.548 0.591 0.577 0.688 0.783 E-9 0.545 0.522 0.426 0.677 0.752 0.830 0.737 0.765  E-10 0.576 0.433 0.595 0.702 0.699 0.768 0.826 0.862  E-11 0.641 0.574 0.819 0.822 0.887 0.890 0.918 0.941 No 0.887 compound

TABLE-US-00014 TABLE 13 Inhibition of growth of Salmonella typhimurium by compounds after 9-20 h: Conc. Absorbance of culture medium at 600 nm (μM): 100 50 25 13 6 3 2 0.8 E-2 0.251 0.168 0.314 0.621 0.382 0.410 0.441 0.452 E-5 0.390 0.395 0.396 0.437 0.478 0.511 0.568 0.574  E-13 0.398 0.407 0.427 0.459 0.505 0.543 0.605 0.634 E-1 0.557 0.484 0.736 0.722 0.741 0.711 0.761 0.722  E-12 0.332 0.270 0.326 0.343 0.400 0.354 0.457 0.455 E-9 0.221 0.319 0.395 0.382 0.348 0.327 0.272 0.300  E-10 0.280 0.315 0.390 0.399 0.382 0.402 0.361 0.334  E-11 0.562 0.650 0.697 0.633 0.627 0.623 0.587 0.555 No 0.943 compound

Example 37

[0434] Substances may act directly on bacteria, or they may act to promote the killing of the bacteria by phagocytes. To measure this effect, cultures murine macrophages are incubated with a test bacteria and the number of bacteria surviving are counted in terms of the viable colony forming units (CFU). The method for determining the rate of phagocytosis is as follows:

[0435] Intracellular killing of S. Typhimurium by mouse macrophage cellline J 774 A.1

[0436] seed a monolayer of cells in 200 μl Media into the wells of a 96 well plates

[0437] incubate O/N 37° C.

[0438] remove medium and add fresh medium

[0439] add Bacteria (Salmonella typhimurium) e.g. 5 μl of 1:100 diluted O/N culture (MOI=10) (=108 cfu/ml)

[0440] centrifuge 10 min 800 g (˜2000 rpm)

[0441] incubate 20-30 minutes at 37° C. (phagocytosis)

[0442] remove media

[0443] wash 1-2× with PBS

[0444] add medium with 100 μg/ml Gentamicin, stock: 10 mg/ml (=1:100)

[0445] incubate 45′ at 37° C.

[0446] wash 2× with PBS

[0447] add fresh medium (200 μl/well)

[0448] add compounds to test

[0449] incubate 2-3 hours at 37° C.

[0450] remove medium

[0451] lyse cells with water: add 200 μl H2O incubate 10′, push a few times through 27 gauge needle using a 1 ml syringe

[0452] plate 100 μlf 1:10 dilution onto LB-agar plates (=1:100 dil)

[0453] Monolayer of J 774 A.1 in 96 well plate=˜1-5×104 cells

[0454] Overnight culture of Salmonella thyph.=˜1×1010 cfu/ml

[0455] Overnight culture of Staph. carnosus=˜5×109 cfu/ml MOI=50 (=5 μl of 1:10 dil. O/N culture)

[0456] MOI=Multiplicity of Infection

[0457] Medium: DMEM/RPMI 7.5 FCS

Example 38

[0458] The potential efficacy of a Compound for Inflammatory bowel disease may be modeled as follows. C57 BLK6 or BALBc mice are provided with drinking water containing 2.5% or 2.8% dextran sulfate. Animals are weighed and observed for signs of intestinal disturbance daily. Signs include diarrhea or occult blood. Compound is formulated by mixing with a solution of 0.1 up to 1% citric acid depending on concentration. Compound is provided by oral gavage daily. Example data for the efficacy of compounds cited here is provided in FIG. 3, 7, 8 or 10-19.

Example 39

[0459] The potential efficacy of a Compound for rheumatoid arthritis may be modeled as follows. DBA1 mice are induced by a subcutaneous injection of bovine collagen in 0.05M acetic acid, emulsified in Freund's adjuvant. 21 days later, a second injection of this material is made without inclusion of mycobacterial material in the adjuvant. Animals are weighed and observed for signs of inflammation daily. Signs include weight loss, swelling of paws, redness and reduced mobility. Compound is formulated by mixing with a solution of 1% citric acid. Compound is provided by oral gavage daily. Data for the efficacy of compounds cited here is provided in FIG. 2.

Example 40

[0460] The potential efficacy of a Compound in modulating immune reactions may be determined as follows. Swiss or C57 Blk6 mice are induced to produce cytokines by a subcutaneous injection of lipopolysaccharide, Typically, compound is provided at time 0. Compound is formulated by mixing with a solution of 1% citric acid for oral treatment or, dissolved in PEG 300 and diluted in water for intra-peritoneal treatment. Compound is provided by oral gavage. 30 minutes after providing compound, animals are treated with an intra-peritoneal injection of a solution of lipopolysaccharide in the concentration range that will provide 0.01 mg/kg lipopolysaccharide. Data for the efficacy of compounds cited here is provided in FIG. 1.

Example 41

[0461] The potential efficacy of a Compound in treating a malignant disease may be determined as follows. Tumours are known to be deficient in nitric oxide and this is considered to be a cause of local tolerance. Providing a nitric oxide donor that is accumulated in macrophages in the tumour environment provides a means to artificially modify the local NO status. C57 Blk6 mice are injected subcutaneously with an murine ovarian cancer cell line expressing ovalbumin. Mice bearing tumours are selected after 14 days. Typically, compound is provided at this time. Compound is formulated by mixing with a solution of 1% citric acid for oral treatment. Compound is provided by oral gavage. Animals are monitored daily for tumour size, body weight and activity score. The activity of the compound may be determined in combination with other therapies including anti-bodies or vaccines based on a tumour antigen. In this case ovalbumin, can serve as a model antigen.

Example 42

[0462] Synthesis of 2′-O-(2-Ferrocenyl) Acetyl-11-O-Nitro-Azithromycin

##STR00094##

[0463] 11-O-Nitro-azithromycin (0.25 mmol) was dissolved in dry dichloromethane (5 mL). To this was added EDCI (2 eq., 0.5 mmol) and 2-ferrocenyl acetic acid (1.1 eq., 0.28 mmol). The reaction was stirred overnight at room temperature. The solvent was removed in vacuo and the resulting white amorphous foam. The resulting crude product was purified by column chromatography with a gradient starting at 10% of acetone in cyclohexane (0.2% Et.sub.3N).

[0464] Similarly, the following compound may be obtained using the procedure above starting from 2′-O-Nitro-azithromycin:

[0465] 2′-O-Nitro-11-O-(2-Ferrocenyl) Acetyl-Azithromycin

##STR00095##

Example 43

[0466] Rac. 2′-O-Propionyl Propranolol

##STR00096##

[0467] Diethyl azodicarboxylat (DEAD, 10 mmol) is added to a stirred (magnetic stirrer, 300 rpm) solution of triphenylphosphin (10 mmol) in dry THF (2.5 mL) at 0° C. and treatment is continued for 30 min. Propranolol (5 mmol) and propionic acid (10 mmol) both dissolved in THF (10 mL) are added dropwise and stirring is continued for 1 h at 0° C. and further 2 hours at ambient temperature. Any precipitates are filtered off, the remaining solution is concentrated in vacuo and desired product is isolated by column chromatography (silica gel, cyclohexane-ethyl acetate).

Example 44

[0468] Rac. 2′-O-Acetoxypropionyl Propranolol

##STR00097##

[0469] Diethyl azodicarboxylate (DEAD, 10 mmol) is added to a stirred (magnetic stirrer, 300 rpm) solution of triphenylphosphin (10 mmol) in dry THF (25 mL) at 0° C. and treatment is continued for 30 min. Propranolol (5 mmol) and 2-acetoxypropionic acid (10 mmol) both dissolved in THF (10 mL) are added dropwise and stirring is continued for 1 h at 0° C. and further 2 hours at ambient temperature. Any precipitates are filtered off, the remaining solution is concentrated in vacuo and desired product is isolated by column chromatography (silica gel, cyclohexane-ethyl acetate).

Example 45

[0470] Synthesis of Azithromycin 11,2′-Lipoate

##STR00098##

[0471] 380 mg of azithromycin-2′-lipoate (E-77) are dissolved in 25 ml of dichloromethane and cooled in an ice bath. 125 mg of lipoyl are adde, then 50 μl of pyridine. The mixture is allowed to reach room temperature and stirred for 16 h. The reaction mixture is extracted with water 3 times, then once with 5% aqueous citric acid. The citric acid phase is extracted with dichloromethane, then combined with ethyl acetate and carefully made basic with sodium hydrogen carbonate and vigorous stirring. When gas evolution ceases, the organic phase is separated, washed with water and brine, and dried with sodium sulfate. After evaporation of all volatiles, the residue is chromatographed with a gradient starting at cyclohexane-acetone 5-1, containing 0.5% of triethylamine. Yield: 140 mg

Example 46

[0472] Synthesis of Azithromycin 11-Lipoate

##STR00099##

[0473] 350 mg of Azithromycin 11,2′-dilipoate are stirred with 5 ml methanol at room temperature. When mass spectrometry indicates completion of the reaction (m/z=1125.5.fwdarw.937.5), the mixture is partitioned between water and ethyl acetate. The organic phase is washed once with water, then extracted with 5% aqueous citric acid. The citric acid phase is extracted with dichloromethane, then combined with ethyl acetate and carefully made basic with sodium hydrogen carbonate and vigorous stirring. When gas evolution ceases, the organic phase is separated, washed with water and brine, and dried with sodium sulfate. After evaporation of all volatiles, the residue is chromatographed with a gradient starting at cyclohexane-acetone 5-1, containing 0.5% of triethylamine. Yield: 225 mg

Example 47

[0474] Formation of Acetic Esters of ALCs

[0475] Method 1: ALC (1.0 mmol) was taken up in 15 mL dichloromethane. Pyridine (1.2 eq.) was added and the resulting solution was cooled in an ice bath for approximately 10 minutes. At this point, a solution of acetic anhydride (1.2 eq) was added dropwise. The reaction was stirred continually at this temperature and then progressively warmed to room temperature where it was stirred overnight. Reaction progress was monitored either by TLC and/or MS. The reaction was washed with a saturated solution of ammonium chloride (3×), water (3×) and dried over anhydrous Na.sub.2SO.sub.4. The solvent was evaporated in vacuo. Co-evaporation with toluene is necessary to remove residual pyridine from the system. This was followed by re-dissolving the residue in DCM and solvent evaporation twice to produce a white foam, which was dried under high-vacuum to produce acetylated product.

[0476] This acetylation conditions can be extended for other ALCs. In the case where the acetylation proceeds sluggish, alternative reaction conditions were undertaken as described below:

[0477] Method 2. Compound A-12 (0.85 mmol) was taken up in DCM (10 mL). To this was added triethylamine (3.5 eq) and acetyl chloride (3.5 eq). Reaction was monitored by TLC and MS until disappearance of starting ALC. Reaction was filtered. The filtrate was either evaporated in vacuo and directly purified by column chromatography or the filtrate was washed with 10% aq. Na.sub.2CO.sub.3 solution, brine, dried over Na.sub.2SO.sub.4 and evaporated in vacuo to get the crude product.

[0478] Method 3. Acetic acid (4 eq) was taken up in 5 mL dichloromethane (DCM). Compound A-16 (0.5 mmol) and 4-dimethylaminopyridine (DMAP) (4.4 eq) were added and the resulting solution was cooled in an ice bath for approximately 10 minutes. At this point, dicyclohexylcarbodiimide (DCC) (4.4 eq) was added slowly. The reaction was stirred continually at this temperature for 5 minutes and then progressively warmed to room temperature where it was stirred overnight. Dicyclohexylurea (DCU) that was formed during the reaction is filtered off and discarded. The filtrate was collected and then washed with a saturated solution of sodium hydrogencarbonate (3×), water (1×) and dried over anhydrous Na.sub.2SO.sub.4. The solvent was evaporated in vacuo. This was followed by re-dissolving the residue in a small volume of methanol. The solution was transported dropwise into ice-cold water (2× volume of methanol) and stored in the freezer overnight. The precipitated product was filtered off and dried under high-vacuum to produce a product.

TABLE-US-00015 TABLE 14 Acetylation Examples Reaction MS Compound Synthesis Substituent Condition Degree of m/z Entry Method ALC equivalent (i.e. Workup) Substitution Yield ([M + H].sup.+) E-2 1 A-1 1.2 as described above 1 76% 837 E-4 1 A-1 1.5 as described above 1 41% E-8 1 A-1 1.2 39%  E-23 1 A-1 2.0 as described above 2 54%  E-25 1 A-1 1.1 as described above 1 67%  E-418 3 A-16 4 as described above 3 20% CHMA02063  E-228 1 A-10 4 as described above 2 56% 673  E-453 A-17 Overall 1 and 2 49% 819.7 A-17 12 equiv. (referring to 861.5 Ac.sub.2O the di-ester)   E-266/ 2 A-12 3 as described above, 2 > 4 77% 675, 759  E-268 filtered through a silica gel plug using CHCl.sub.3:iPrOH:7M NH.sub.3 in MeOH (30:1:1) as mobile phase E-23/E-50 A-1 2 and 3 16% 833, 875  E-564 1 A-2 (X = O) 1 89% 776  E-29 1 E-19 1.2 direct chromatography 1× 85% 861 for purification

Example 48

[0479] Formation of Butyric and Isobutyric Esters of ALCs

[0480] Method 1: Compound A-1 was taken up in dichloromethane and stirred for 10 min. At this point, a solution of carboxylic anhydride and triethylamine in dichloromethane was added dropwise. The reaction was stirred continually at room temperature. The reaction solution was washed with 5% citric acid three times to extract the product. Acidic solution was then washed with ethyl acetate (2×) and afterwards neutralized with Na.sub.2CO.sub.3. Product was extracted with ethyl acetate (3×). The solution was washed with a saturated solution of sodium chloride (2×), water (2×) and dried over anhydrous Na.sub.2SO.sub.4. The to solvent was evaporated in vacuo to produce a white foam containing product.

[0481] Method 2: Compound A-1 was taken up in dichloromethane and was cooled in an ice bath for approximately 10 minutes. At this point, a solution of carboxylic chloride in dichloromethane was added dropwise. The reaction was stirred continually at this temperature for 15 min and then progressively warmed to room temperature where it was stirred for 2.5 h.

[0482] The reaction was washed with a 10% solution of Na.sub.2CO.sub.3 (3×), water (3×) and dried over anhydrous Na.sub.2SO.sub.4. The solvent was evaporated in vacuo. Co-evaporation with toluene is necessary three times. This was followed by re-dissolving the residue in dichloromethane to produce a white foam, which was dried under high-vacuum to produce product.

[0483] Method 3: Starting material was taken up in dichloromethane and stirred for 10 min. At this point, a solution of carboxylic chloride and triethylamine in dichloromethane was added dropwise. The reaction was stirred continually at room temperature for two days. The reaction solution was washed with 5% citric acid three times to extract the product. Acidic solution was then washed with ethyl acetate (2×) and afterwards neutralized with Na.sub.2CO.sub.3. Product was extracted with ethyl acetate (3×). The solution was washed with a saturated solution of sodium chloride (2×), water (2×) and dried over anhydrous Na.sub.2SO.sub.4. The solvent was evaporated in vacuo to produce a white foam containing product.

[0484] Method 4: Compound E-48 or E-39 was solved in methanol to hydrolyze butyric esters. The reaction was stirred continually at room temperature for two days. The reaction solution was washed with ethyl acetate three times to extract the product. The ethyl acetate phase was washed with 5% citric acid (3×). Acidic solution was then washed with ethyl acetate (2×) and afterwards neutralized with Na.sub.2CO.sub.3. Product was extracted with ethyl acetate (3×). The solution was washed with a saturated solution of sodium chloride (2×), water (2×) and dried over anhydrous Na.sub.2SO.sub.4. The solvent was evaporated in vacuo to produce a white foam containing product.

[0485] Method 5: Carboxylic acid (180 mg, 2.04 mmol) was solved in 3 mL dichloromethane. Under stirring conditions 4-Dimethylaminopyridine (274 mg, 2.24 mmol) and A-16 were added. The reaction solution was cooled to 0° C. and N,N′-Dicyclohexylcarbodiimide (463 mg, 2.24 mmol) was added. The reaction was stirred continually at this temperature for 5 min and then progressively warmed to room temperature where it was stirred for 12 h. Precipitation was removed via filtration. The reaction was washed with a saturated solution of NaHCO.sub.3 (3×) and dried over anhydrous Na.sub.2SO.sub.4. The solvent was evaporated in vacuo. Product was solved in methanol and water was added. Solution was cooled to −20° C., a precipitation occurred which was extracted and dried in vacuo.

[0486] Workup 1: Column chromatography over silica gel was carried out to separate different products. As eluent a mixture of chloroform, 2-propanol and ammonia in methanol (60:1:1) was used. The solvent was evaporated in vacuo.

[0487] Workup 2: Preparative chromatography over RP-C18-silica gel was carried out to separate different products. As eluent a mixture of water (with trifluoroacetic acid 0.05%) and methanol (with trifluoroacetic acid 0.05%) was used. The solvent was evaporated in vacuo.

[0488] Workup 3: Column chromatography over silica gel was carried out to separate different products. As eluent a mixture of cyclohexane, acetone (7:1) with 0.5% triethylamine was used. The solvent was evaporated in vacuo.

TABLE-US-00016 TABLE 15 Butyrylation/Isobutyrylation Examples Compound Synthesis Reaction Condition Degree of Entry Method ALC Substituent (i.e. Workup) Substitution Yield MS E-35 1 A-1 Butyric Workup 1 1x 76% 819 E-39 3 A-1 Butyric Workup 2 2x 10% 889 E-48 3 A-1 Butyric Workup 2 3x 12% 959 E-47 3 A-1 Butyric Workup 2 4x 10% 1029   E-424 5  A-16 Butyric none 3x 97% 944  E-458 2  A-17 Butyric none 1x 89% 847 E-19 4  E-39 Butyric Workup 2 1x 85% 819 E-82 4  E-48 Butyric Workup 3 1x 35% 819  E-553 4  E-48 Butyric Workup 3 2x 30% 889 E-44 1 A-1 Isobutyric Workup 1 1x 67% 819   E-458, 3  A-17 Butyric Workup 1 1x, 2x, 3x 86%  847,   E-459,  917,  E-460 987  E-238 1  A-10 Butyric none 1x 96% 659   E-241, 3  A-10 Butyric Workup 1 2x, 3x, 4x 85%  730,   E-249, 800, 870  E-250  E-111 3 E-1 Butyric Workup 2 1x 38% 864  E-255* 1  A-11 Butyric Workup 1 2x  3% 892  E-256* 1  A-11 Butyric Workup 1 3x 5.6%  964  E-85* 1  A-11 Butyric Workup 1 3x 964  E-257* 1  A-11 Butyric Workup 1 4x  3% 1036  E-24 3 E-1 Butyric Workup 1 1x 75 864 E-89 1 A-3 Butyric n/a 1x  9% 406 *isolated from one reaction

Example 49

[0489] Formation of Valeric Esters of ALCs

[0490] Method 1. The ALC was taken up in DCM. To this were added pyridine and valeric acid anhydride (1 equiv. pyridine/1 equiv. valeric acid anhydride). The mixture was stirred at room overnight or over the weekend and was then poured on an aqueous citric acid solution (5% or 10%; at RT and was stirred for 15 min. The aqueous phase was extracted with EtOAc (2×) and was afterwards brought to pH=8 with solid Na.sub.2CO.sub.3. The alkaline aqueous layer was extracted with EtOAc (2×) and the combined organic phases were washed with water (1×) and saturated aqueous NaCl-solution (1×), dried (Na.sub.2SO.sub.4), concentrated to dryness and dried at the oil pump. Products were obtained as colorless solids or foams.

[0491] Method 2. Analogue to 2A but after stirring at RT for 2 h additional pyridine (2 equiv.) and valeric acid anhydride (2 equiv.) were added and stirring was continued overnight. Products were obtained as colorless foams or solids.

[0492] Method 3. The ALC was taken up in DCM. To this were added pyridine (4 equiv.) and valeric acid anhydride (4 equiv.). The mixture was stirred at room overnight or over the weekend. Additional pyridine (2 equiv.) and valeric acid anhydride (2 equiv.) were added and the mixture was stirred overnight. Reaction mixture was filled into a separation funnel and washed with saturated aqueous NH.sub.4Cl-solution (3×) and water (3×). The organic phase was dried (Na.sub.2SO.sub.4) and concentrated to dryness. The residue was co-evaporated with toluene (3×) and with DCM (3×). Afterwards the crude product was purified by column chromatography on silica gel. Eluent: Chloroform/Isopropanol/NH.sub.3 (7 M in Methanol) 30/1/1

[0493] The product was dried at the oil pump. Products were obtained as colorless solids or foams.

TABLE-US-00017 TABLE 16 Valeric Ester Examples Overall MS Compound Synthesis equiv. of acid Degree of m/z Entry Method ALC or anhydride Re Substitution Yield ([M + H].sup.+) E-72  3 A-1 6 Amount DCM: 5 mL 2 65% 917.5 E-558 1 A 2 5 Anhydro erythromycin 1  7% 800.5 (Anhydro) products are formed Amount DCM: 25 mL E-569 3  A-10 6 Amount DCM: 5 mL 2 45% 757.5 E-582 2  A-12 6 Amount DCM: 10 mL 1 89% 675.5 Citric acid: 10% E-411 1  A-15 4 Amount DCM; 25 mL 1 and 2 81% 878.3 E-412 Citric acid: 5% 962.3 E-428 1  A-16 3 Amount DCM: 25 mL 1 and 2 83% 819.0 E-429 Citric acid: 5% 902.8 E-464 1 A17 5 Amount DCM: 25 mL 1 94% 861.7 Citric acid: 5%

Example 50

[0494] Formation of Isovaleric Esters of ALCs

[0495] Method 1: Isovaleric acid (4.4 equiv./equiv. ALC) and HOBt 85% (4.4 equiv./equiv. ALC) were dissolved in DMF (12.5 mml/mmol ALC). The solution Was cooled down to 0-5° C. in an ice-bath. At this temperature a solution of Dicyclohexylcarbodiimide (4.5 equiv./equiv. ALC) in DCM (5 ml/mmol ALC) was added dropwise within 30 min. the solution was kept at this temperature for another 10 min. Then Azithromycin (1 equiv.) was added in one portion. While stirring, the solution was allowed to come to room temperature within 2 h. Stirring was continued for another 2 h at 50° C. The reaction mixture was allowed to stand at RT for 12 h. A white precipitate was removed by suction. The solvent was evaporated completely at 12 mbar and 50° C. The residue was dissolved in DCM (12.5 mL/mmol ALC) and washed with water (7.5/mmol ALC). A small amount of a white precipitate was removed. Then the solution was treated with citric acid (25 mL/mmol ALC, 5%). The aqueous phase was washed with DCM (5 ml/mmol ALC). NaOH 10% was added until the aqueous phase was basic (pH 12) and was washed with DCM (2×10 ml/mmol ALC). After phase separation the organic phase was evaporated to dryness, products were obtained as white solids.

[0496] Method 2A. The ALC was taken up in DCM. To this were added pyridine and isovaleric acid anhydride (1 equiv. pyridine/1 equiv. isovaleric acid anhydride). The mixture was stirred at room overnight or over the weekend and was then poured on an aqueous citric acid solution (5% or 10%) at RT and was stirred for 15 min. The aqueous phase was extracted with EtOAc (2×) and was afterwards brought to pH=8 with solid Na.sub.2CO.sub.3. The alkaline aqueous layer was extracted with EtOAc (2×) and the combined organic phases were washed with water (1×) and saturated aqueous NaCl-solution (1×), dried (Na.sub.2SO.sub.4), concentrated to dryness and dried at the oil pump. Products were obtained as colorless solids or foams.

[0497] Method 2B. Analogue to 2A but after stirring at room temperature for 2 h additional pyridine (2 equiv.) and isovaleric acid anhydride (2 equiv.) were added and stirring was continued overnight.

[0498] Products were obtained as colorless foams or solids.

[0499] Method 2C. The ALC was taken up in DCM. To this were added pyridine (4 equiv.) and isovaleric acid anhydride (4 equiv.). The mixture was stirred at room for approximately 2 h, then a catalytic amount of DMAP was added, followed by another catalytic amount approximately another 2 h later. The mixture was stirred at room temperature for approximately 2 h before additional pyridine (2 equiv.) and isovaleric acid anhydride (2 equiv.) were added. The mixture was stirred at room overnight and then poured on an aqueous citric acid solution (5%,) and stirred at room temperature for 30 min. The aqueous phase was extracted with EtOAc and afterwards brought to pH=8 with solid Na.sub.2CO.sub.3. The alkaline aqueous layer was extracted with EtOAc (2×) and the combined organic phases were washed with water (1×) and saturated aqueous NaCl-solution (1×), dried (Na.sub.2SO.sub.4), concentrated to dryness and dried at the oil pump. Products were obtained as colorless solids or foams.

TABLE-US-00018 TABLE 17 Isovaleric Ester Examples Overall MS Compound Synthesis equiv. of acid Degree of m/z Entry Method ALC or anhydride Annotations Substitution Yield ([M + H].sup.+) E-45  1  A-1 1 91% 833.5 E-557 2A A 2 5 Anhydro erythromycin 1 17% 800.5 (Anhydro) products are formed Amount DCM: 25 mL Citric acid: 5% E-573 2C A-10 6 Amount DCM: 7.5 mL 2 19% 757.5 Citric acid: 5% E-583 28 A12  6 Amount DCM: 10 mL 1 83% 675.5 Citric acid: 10% E-413 2A A-15 4 Amount DCM: 25 mL 1 95% 878.3 Citric acid: 5% E-431 2A A-16 3 Amount DCM: 25 mL 2 87% 818.8 E-432 Citric acid: 5% 902.7 E-467 2A A-17 5 Amount DCM: 25 mL 1 90% 861.6 Citric acid: 5%

Example 51

[0500] Long Chain (>C5) Fatty Acid Substation of Tildipirosin

[0501] Hexanoic acid (290 mg, 2.5 mmol) was taken up in 5 mL dichloromethane (DCM). Compound A-16 (367 mg, 0.5 mmol) and 4-Dimethylaminopyridine (DMAP) (336 mg, 2.75 mmol) were added and the resulting solution was cooled in an ice bath for approximately 10 minutes. At this point, dicyclohexylcarbodiimide (DCC) (567 mg, 2.75 mmol) was added slowly. The reaction was stirred continually at this temperature for 5 minutes and then progressively warmed to room temperature where it was stirred overnight. Dicyclohexylurea (DCU) that was formed during the reaction is filtered off and discarded. The filtrate was collected and then washed with a saturated solution of ammonium chloride (3×), sodium hydrogencarbonate (3×), water (1×) and dried over anhydrous Na.sub.2SO.sub.4. The solvent was evaporated in vacuo. This was followed by re-dissolving the residue in a small volume of methanol. The solution was transported dropwise into ice-cold water (2× volume of methanol) and stored in the freezer overnight. The precipitated product was filtered off and dried under high-vacuum to produce a mixture of E-437, E-438 and E-439 (56%).

[0502] This esterification conditions can be extended for other acids.

TABLE-US-00019 TABLE 18 Long chain fatty acid substitution of ALC Degree of Compound Substituent Substitu- Entry Acid equivalent tion Yield MS E-437, Hexanoic acid 5 2 and 3 56% 931.7 E-438, 1028.8 E-439 E-440, Heptanoic acid 5 1, 2 and 3 46% 846.9 E-441, 959.3 E-442 1071.9 E-445 Octanoic acid 5 3 17% 1113.5 E-447, Decanoic acid 5 2 and 3 41% 1043.3 E-448 1197.5 E-450, Dodecanoic 5 2 and 3 60% 1099.5 E-451 acid 1281.4

Example 52

[0503] General Procedure for Preparing Cores A-13 and A-14

##STR00100##

[0504] Macrolide (1 mmol) is dissolved in DMF (500 μl). Epichlorohydrin (1 mL) is added and the mixture is heated to 80° C. for 12 h. When MS analysis indicates complete conversion, all volatiles are removed in vacuo and the residue is dissolved in ethanol (1 ml). The solution is poured into 25 ml of water. The precipitate is isolated and can be used directly for the next step or is chromatographed to obtain the pure epoxide.

[0505] The following macrolides were used to form epoxides as precursor to the desired cores.

TABLE-US-00020 TABLE 19 Epoxide Formation MW Entry Macrolide epoxide Yield 1 Azithromycin 703 25% 2 Gamithromycin 731 12% 3 3-decladinosyl-3-oxoazithromycin 543  5% 4 Tildipirosin 689  5%

[0506] Epoxide (1 mmol) is dissolved in 2-propanol (500 μl), and an excess of 5 equivalent of an amine is added. The mixture is heated from 12 h to 100 h at 80° C. When MS indicates complete conversion, all volatiles are evaporated and the residue subjected to chromatography to separate the 2 regioisomeric amines.

TABLE-US-00021 TABLE 20 Epoxide opening by amines Epoxide Entry opening (Table 9) Amine position R.sup.1 R.sup.2 [M + H].sup.+ Yield [%] Comment 1 dimethylamine 3′ Me Me 749 66 NMR identical to azithromycin 2′ 22 regioisomer of Azithromycin 1 morpholine 3′ R.sup.1 = R.sup.2 = morpholine ring 791 30 unpolar product (A-13.1) 2′ 61 polar product (A-13.2) 1 diethanolamine 3′ C.sub.2H.sub.4OH C.sub.2H.sub.4OH 809 22 unpolar product (A-14.1) 2′ 47 polar product (A-14.2) 1 ammonia H H 721 n.d.* 1 N-methyl Me OH 751 n.d.* hydroxylamine 1 Iminodiacetic —CH.sub.2C(O)OEt —CH.sub.2C(O)OEt 893 n.d.* some cyclized acid product with diethylester [M + H].sup.+ = 865 is formed, too 2 morpholine R.sup.1 = R.sup.2 = morpholine ring n.d.* *n.d. not determined

Example 53

[0507] Further Acylation Reactions of ALC

[0508] Method 1: Compound A-1 (2000 mg, 2.67 mmol) was taken up in 10 mL dichloromethane and stirred. Separately 4.4 eq of a carboxylic acid and 4.4 eq of 1,1′-Carbonyldiimidazole were solved in dichloromethane (10 mL) and stirred over 20 min. Both solutions were unified and stirred continually at room temperature. The dichloromethane phase was washed with saturated NaHCO.sub.3 solution (2×) and dried with Na.sub.2SO.sub.4 (anhydrous). The solvent was evaporated in vacuo to produce a white foam containing products of reaction.

[0509] Method 2: Compound E-1 (265 mg, 0.33 mmol) was taken up in 10 mL dichloromethane and stirred. Separately 1.6 eq of methoxyacetic acid and 1.6 eq of 1,1′-Carbonyldiimidazole were solved in dichloromethane (5 mL) and stirred over 20 min. Both solutions were unified and stirred continually at room temperature. The reaction solution was washed with 5% citric acid three times to extract the product. Acidic solution was then washed with ethyl acetate (2×) and afterwards neutralized with Na.sub.2CO.sub.3. Product was extracted with ethyl acetate (3×). The solution was washed with a saturated solution of sodium chloride (2×), water (2×) and dried over anhydrous Na.sub.2SO.sub.4. The solvent was evaporated in vacuo to produce a white foam containing E12 (238 mg, 90%).

[0510] Workup 1: Column chromatography over silica gel was carried out to separate different products. As eluent a mixture of chloroform, 2-propanol and ammonia in methanol (60:1:1) was used. The solvent was evaporated in vacuo.

[0511] Workup 2: Column chromatography over silica gel was carried out to separate different products. As eluent a mixture of cyclohexane, acetone (3:1) with 0.5% triethylamine was used. The solvent was evaporated in vacuo.

TABLE-US-00022 TABLE 21 Typical Products from the Acylation Procedures (2) Reaction Cpd Synthesis Condition Degree of Entry Method Carboxylic acid ALC (i.e. Workup) Substitution Yield MS E-545 1 Cyclopropanecarboxylic acid A-1 Workup 1 1×   30% 817 E-546 1 Cyclobutanecarboxylic acid A-1 Workup 1 1×   10% 830  E-546, 1 Cyclobutanecarboxylic acid A-1 Workup 1 1×, 2×, 3× 22.5%  830,  E-560,  913, E-561 995 E-547 1 Nicotinic acid A-1 Workup 2 1× 10.3% 854  E-547, 1 Nicotinic acid A-1 Workup 2 1×, 2× 11.8%  854, E-562 959 E-551 1 Methoxyacetic acid A-1 Workup 1 1× 4.4 820 E-14,  1 Methoxyacetic acid A-1 2×, 3×, 4× Reaction  893, E-22,  solution  965, E-559 1037  E-12  2 Methoxyacetic acid E-1 none 1×   90% 866 E-81  1 3-Phenylpropionic acid A-1 Workup 1 1×   30% 881 E-544  1* Indole-3-propionic acid A-1 Workup 1 1×   45% 920 *instead of 1,1′-carbonyl diimidazole, HATU was used as coupling agent.

Example 54

[0512] Synthesis of E-541 and E-542

##STR00101##

[0513] E-27 (1.2 mmol) and Quinoline-amine (1 eq) was taken up in DCM (5 mL). To this was added HATU (1.2 eq) neat. The reaction was stirred overnight at room temperature. Reaction was very sluggish an additional 0.5 eq of HATU was added. Reaction was stirred for 2 days or disappearance of starting material was observed (TLC or MS). Reaction solution was removed in vacuo and the crude material directly purified by chromatography to get the desired product.

TABLE-US-00023 TABLE 22 Decoration of ALC (E-27) with potential TLR-agents Cpd Degree of Entry Quinoline Substitution Yield MS E-541 Imiquimod 1 35% 1071 E-542 Resiquimod 1 25% 1145

Example 55

[0514] Examples of Polyamines as ALC

[0515] These ALCs can be prepared by reacting symmetrical or unsymmetrical di- or poly-epoxides with secondary amines. This will provide ALCs that contains common structural element of 2 or more alcohols, vicinally neighbored by a tertiary amine. Some polyamines are also commercially available.

[0516] Alternatively, ALCs can be prepared by reacting epoxides with diethanolamine. The reaction products are containing 2-hydroxy tertiary amines.

[0517] General Procedure for Preparation of some polyamine ALC Bearing hydroxy Functionalities:

[0518] Polyamine (1 mmol) containing at least 2 NH-functions and the epoxide are mixed and heated without solvent to 80° C. Excess epoxide can be removed by column chromatography selectively. Products are sufficient, when at least 2 tertiary ß-hydroxyamines are present.

TABLE-US-00024 TABLE 23 Examples of Polyamine ALCs Eq. of Reaction [M + H].sup.+ Entry Amine Epoxide Epoxide time Product Yield Remark 1 hexamethylene diamine 1,2- 4.2  40 h 966 n.d. product contains epoxytetradecane a small amount of triple alkylation 2 bis- cyclohexene 12 240 h 608 n.d. main product is (hexamethylene)triamine oxide tetraalkylated

##STR00102##

[0519] Reactions of Polyepoxides with Amines:

[0520] Corresponding polyepoxide (1 mmol) is mixed with 1.05 mmol secondary amine per epoxide function and heated to 80° C. without solvent for 12 h.

TABLE-US-00025 TABLE 24 Further Examples Polyamine ALCs [M + H].sup.+ Entry Epoxide Amine Product Yield 1 1,2,7,8-diepoxyoctane morpholine 317 100% 2 Diglycidyl ethylenglycol morpholine 377  90%

##STR00103##

[0521] Synthesis of E-552

##STR00104##

[0522] 1.45 g of 1,10-dimorpholino-2,9-dihydoxy-4,7-dioxadecane are combined with 1.5 ml of butyric anhydride in 5 ml of chloroform. After stirring for 1 h, MS indicates complete conversion ([M+H].sup.+=517). The mixture is extracted with 2 N KOH, saturated aqueous sodium bicarbonate solution and brine, dried and chromatographed over silica gel to obtain 1.57 g of the target compound (72%).

Example 56

[0523] Substitution of Polyamine ALC (see Table 1, Entry A-19)

[0524] Substitution ALC A-19.1/A-20.1/A-20.2

[0525] Method 1: N-Hydroxyalkyl compound (5 mmol) was suspended in excess carboxyl acid anhydride (>100 mmol, >20 eq.) in a round bottom flask while stirring (magnetic stir bar, 500 rpm). The mixture was cooled in an ice bath and sulfuric acid (>96%, 3 drops) was carefully added as catalyst. Stirring was continued until a clear solution was obtained. When ESI-MS indicated full conversion of starting materials the reaction mixture was poured on ice. The system was stirred for 2 or more hours in order to hydrolyze any anhydride. The mixture was neutralized by addition of sodium bicarbonate and extracted with dichloromethane (3×). Separation of organic phase, drying over sodium sulfate and evaporation of any volatiles in vacuo yielded the product as colorless oil.

[0526] Method 2. Carboxylic acid (1.5 eq per hydroxyl group) was placed into a round bottom flask along with a stir bar and carbonyl diimidazole (CDI, 1.6 eq per hydroxyl group). Dichloromethane (DCM, 10 mL per gram carboxylic acid) was added carefully while stirring (500 rpm) at ambient temperature. Immediate formation of carbon dioxide indicated conversion of corresponding acid to the acyl donor (caution: too quick CO.sub.2 formation may result in strong foaming. Do not seal the flask!). After a couple of minutes, a clear solution was obtained and stirring was continued for 15 minutes. N-Hydroxyalkyl compound (5 mmol) was added at ambient temperature and the reaction mixture was stirred overnight. When ESI-MS indicated full conversion of starting materials the reaction was quenched by addition of methanol, converting excess acyl donor to methyl ester. The system was diluted by addition of further DCM and was subject to extraction with saturated sodium bicarbonate solution (3×). Separation of organic phase, drying over sodium sulfate and evaporation of any volatiles in vacuo yielded the product as colorless oil.

TABLE-US-00026 TABLE 25 Substitution of ALC A-19.1/A-20.1/A-20.2 Compound Synthesis Degree of MS Entry Method ALC Substitution Yield [M + H.sup.+ E-524 1 A-19.1 Di acyl 85% 259, M + H.sup.+ E-576 1 A-19.1 Mono acyl 23% 217, M + H.sup.+ E-525 1 A-19.1 Di acyl 94% 287, M + H.sup.+ E-577 1 A-19.1 Mono acyl 12% 231, M + H.sup.+ E-526 2 A-19.1 Di acyl 87% 315, M + H.sup.+ E-578 2 A-19.1 Mono acyl 45% 245, M + H.sup.+ E-527 1 A-19.1 Di acyl 56% 315, M + H.sup.+ E-529 1 A-19.1 Di acyl  8% 343, M + H.sup.+ E-530 2 A-19.1 Di acyl 76% 499, M + H.sup.+ E-538 1 A-20.1 Tri acyl 85% 299, M + Na.sup.+ E-537 1 A-20.1 Tri acyl 76% 383, M + Na.sup.+ E-580 1 A-20.2 Tri acyl 61% 391, M + Na.sup.+; 368, M.sup.−

[0527] Substitution ALC A-19.2

[0528] 1,1′-Carbonyldiimidazole was dissolved in dichloromethane (dry, 25 mL) and to this was added the carboxylic acid slowly at room temperature. The solution was stirred at room temperature before a suspension of N;N,N′,N′-tetrakis (2-hydroxyethyl)-ethylendiamine (A-19.2) in dichloromethane (dry, 5 mL) was added in one portion at room temperature and the mixture was stirred at RT. The reaction mixture was filled into a separation funnel and washed. The organic phase was dried (Na.sub.2SO.sub.4) and concentrated to dryness in vacuo. The crude product was purified by column chromatography.

[0529] Eluent: CHCl.sub.3: Isopropanol: NH.sub.3 (7 M in MeOH)=60:1:1.

TABLE-US-00027 TABLE 26 Substitution of ALC A-19.12 Amount Time for Carboxylic stirring Cpd Amount acid Amount carboxylic Reaction Washing Entry ALC (A-19.2) (mg) CDI acid and CDI Time steps Yield E-531 A-19.2 521 mg Glacial acetic 2.49 g 35 min 22h 30 min NaHCO.sub.3 297 mg (73.4%) acid 15.35 (2 × 20 (45%) 1.64 800 μL mmol mL) mmol 13.99 mmol E-532 A-19.2 498 mg Propionic acid 2.46 g 20 min 1.5 h with Water 144 mg (73.4%) 943 μL 15.17 with argon argon stream (1 × 20 (20%) 1.55 12.6 mmol mmol stream 69 h 45 min mL) mmol under argon sat. atmosphere NaHCO.sub.3 (2 × 20 mL) E-533 A-19.2 495 mg Butyric acid 2.51 g 35 min 46 h 45 min NaHCO.sub.3 20 mg (73.4%) 1.2 mL 15.45 (2 × 20 (pure) 1.54 13.55 mmol mmol mL) 224 mg mmol (with impuri- Ties Overall yield: (19%)

[0530] Analytical Data:

[0531] E-531:

##STR00105##

[0532] ESI-MS (positive): m/z=405.2 [M+H].sup.+, 427.1 [M+Na].sup.+

[0533] Purity according to HPLC (ELSD): >99.9%

[0534] .sup.1H-NMR (300 MHz, CDCl.sub.3): 1.98 (s, 12 H, 4-H, 4′-H, 4″-H, 4″′-H), 2.57 (s, 4 H, 1-H, 1′H), 2.72 (t, J.sub.2,3 and J.sub.2′,3′, J.sub.2″,3″, J.sub.2″′,3″′=6.04, 8 H, 2-H, 2′-H, 2″-H, 2″′-H), 2.72 (t, J.sub.3,2 and J.sub.3′,2′, J.sub.3″,2″, J.sub.3″′,2″′=4.04, 8 H, 2-H, 2′-H, 2″-H, 2″′-H).

[0535] .sup.13C-NMR (75 MHz, CDCl.sub.3): 20.77 (q, C-4, C-4′, C-4″, C-4″′), 53.15 (t, C-2, C-2′, C-2″, C-2″′), 53.44 (t, C-1, C-1′), 62.43 (t, C-3, C-3′, C-3″, C-3″′), 170.74 (s, 4×C=O).

[0536] E-532:

##STR00106##

[0537] ESI-MS (positive): m/z=461.2 [M+H].sup.+, 483.3 [M+Na].sup.+

[0538] Purity according to HPLC (ELSD): >99.9%

[0539] .sup.1H-NMR (300 MHz, CDCl.sub.3): 1.08 (t, J.sub.5,4, J.sub.5′,4′, J.sub.5″,4″, J.sub.5″′,4″′=7.6 Hz, 12 H, 5-H, 5′-H, 5″-H, 5″40 -H), 2.27 (q, J.sub.4,5, J.sub.4′,5′, J.sub.4″,5″, J.sub.4″′,5″′=7.6 Hz), 2.57, 4-H, 4′-H, 4″-H, 4 ″′ 2.60 (s, 4 H, 1-H, 1′H), 2.74 (t, J.sub.2,3 and J.sub.2′,3′, J.sub.2″,3″, J.sub.2″′,3″′=6.04, 8 H, 2-H, 2′-H, 241 -H, 2″′-H), 4.07 (t, J.sub.3,2 and J.sub.3′,2′, J.sub.3″,2″, J.sub.3″′=6.04, 8 H, 2-H, 2′-H, 2″-H, 2″′-H).

[0540] .sup.13C-NMR (75 MHz, CDCl.sub.3): 8.96 (q, C-5, C-5′, C-5″, C-5″′), 27.44 (t, C-4, C-4′, C-4″, C-4″′), 53.22 (t, C-2, C-2′, C-2″, C-2″′), 53.54 (t, C-1, C-1′), 62.36 (t, C-3, C-3′, C-3″, C-3″′), 172.20 (s, 4×C=O).

[0541] E-533:

##STR00107##

[0542] ESI-MS (positive): m/z=617.3 [M+H].sup.+, 539.3 [M+Na].sup.+

[0543] Purity according to HPLC (ELSD): >99.9%

[0544] .sup.13C-NMR (75 MHz, CDCl.sub.3): 13.53 (q, C-6, C-6′, C-6″, C-6″′) 18.26 (t, C-5, C-5′, C-5″, C-5″′), 36.00 (t, C-4, C-4′, C-4″, C-4″′), 53.21 (t, C-2, C-2′, C-2″, C-2″′), 53.45 (t, C-1, C-1′), 62.24 (t, C-3, C-3′, C-3″, C-3″′), 173.36 (s, 4×C=O):

[0545] Substitution ALC A-19.3

[0546] H-L-orn(Boc)2CT Resin (0.68 mmol/g, 100-200 mesh, 2.99 g, 2.07 mmol) was filled into a 20 mL syringe with frit. Dichloromethane (dry, 10 mL), MeOH (2 mL) and diisopropylethylamine (2 mL) are added to the resin for endcapping. The mixture was shaken at room temperature for 30 min, then the liquid was sucked off and the resin was washed (3× dimethylformamide 15 mL, 1× diethylether 15 mL).

[0547] The resin was filled into a 100 mL round bottom flask. DMF (25 mL) was added and the resin was swollen for 5 min. Then diisopropylethylamine (3.8 mL, 22.3 mmol) and 2-bromoethanol (1.434 mL, 20.3 mmol) were added subsequently at room temperature. The reaction mixture was stirred at 60° C. (bath temperature) for 24 h.

[0548] The resin was filled into a 20 mL syringe with frit and was washed:

[0549] 4× dimethylformamide (20 mL), 3× methanol (20 mL), 3× dichloromethane (20 mL), 3× diethyl ether (20 mL).

[0550] Half of the resin (1.035 mmol) was filled into a 20 mL syringe with frit.

[0551] Valeric acid (568 μL, 5.69 mmol) was added to a mixture of dimethylformamide/dichloro-methane 1:1 (10 mL). HOBt*H.sub.2O (870 mg, 5.69 mmol) was added and mixture was stirred at room temperature for 5 min before diisipropylcarbodiimide (881 μL, 5.69 mmol) was added. Stirring at room temperature was continued for 10 min, then the whole mixture was added to the resin and the resin was shaken at room temperature for 5 h.

[0552] The liquid was sucked off and the resin was washed:

[0553] 4× dimethylformamide (10 mL), 3× methanol (10 mL), 3× dichloromethane (10 mL), 3× diethyl ether (10 mL).

[0554] A test cleavage showed the product by mass spectrometry

[0555] ESI-MS (positive): m/z=261.1 [M+H].sup.+

Example 57

[0556] Synthesis of 2′-O-(2-Ferrocenyl) Acetyl-Azithromycin E-549

##STR00108##

[0557] ALC A-1 (0.25 mmol) was dissolved in dry dichloromethane (5 mL). To this was added EDCI (2 eq., 0.5 mmol) and 2-ferrocenyl acetic acid (1.1 eq., 0.28 mmol). The reaction was stirred overnight at room temperature. The solvent was removed in vacuo and the resulting white amorphous foam. The resulting crude product was purified by column chromatography with a gradient starting at 10% of acetone in cyclohexane (0.2% Et.sub.3N).

Example 58

[0558] Synthesis of E-258

[0559] Compound A-11/E-16 was dissolved in dry dichloromethane (DCM) in a round bottom flask equipped with magnetic stir bar. Penta-O-acetyl-α-D-mannopyranoside (1.2 eq) was added and the system was cooled in an ice bath while stirring (300 rpm). Catalytic amount of boron trifluoride diethyl ether complex was carefully added dropwise and the system was allowed to warm up while stirring overnight. Upon dilution with further DCM the mixture was subject to extraction with saturated sodium bicarbonate solution (3×). Separation of organic phase, drying over sodium sulfate and evaporation of any volatiles in vacuo yielded the product as colorless oil or beige to off-white foam. [M+H].sup.+ m/z 907

[0560] The reaction conditions also produced the des-cladinosyl product E-600.

Example 59

[0561] Synthesis of E-550

[0562] 350 mg of compound E 77 are dissolved with 15 ml of carbon disulfide. 250 mg of sulfur are added and the mixture is stirred for 7 days. The mixture is extracted with 5% aqueous citric acid solution. The aqueous extract is combined with 10 ml of ethyl acetate and made alkaline by addition of sodium carbonate with intense stirring. The organic phase is separated, washed with brine, dried over sodium sulfate and concentrated in vacuo to yield 280 mg of a product, that contains various higher sulfides along with some starting material, as indicated by mass spectrometry ([M+H].sup.+=969, 1001, 1033, 1065).

Example 60

[0563] Pharmacokinetics

[0564] The distribution of compounds to target organs is of specific importance to the efficacy of anti-infective compounds. To determine distribution the compounds are formulated and administered to a suitable animal model. Compounds were administered p.o. 10 mg/kg in 2% citric acid in BALBc and organs were recovered at 6 h. Organs were extracted in Acetonitrile (6× volume of the sample), centrifuged at 14000 g for 5 minutes. Samples were analysed by LCMSMS (SCIEX 4500). Data are the mean of 3 animals.

Example 61

[0565] Selection of ALC via Concentration into Immune Cells

[0566] The distribution of compounds to target cells is of specific importance to the efficacy of anti-infective compounds. To determine uptake the compounds are dissolved in DMSO or citric acid and mixed with whole blood, plasma or cell medium. To these solutions are added cultured macrophages, cultured immune cells, bone marrow derived macrophages, peritoneal macrophages or buffy coat cells. The mixture is incubated at 37° C. for 1, 2, or 3 hours. After incubation, the immune cells are separated from the medium and the concentration of the compounds is determined by extraction in Acetonitrile (6× volume of the sample), followed by centrifugation at 14000 g for 5 minutes. The resulting extracts are analyzed by LCMSMS (SCIEX 4500 in positive mode). Data are the mean of 3 animals.

[0567] Other Embodiments

[0568] All of the features disclosed in this specification may be combined in any combination. Thus, unless expressly stated otherwise, each feature disclosed is only an example of a generic series of equivalent or similar features.

[0569] It is to be understood that while the invention has been described in conjunction with the detailed description thereof, the foregoing description is intended to illustrate and not limit the scope of the invention, which is defined by the scope of the appended claims. Other aspects, advantages, and modifications are within the scope of the following claims.

[0570] Abbreviations

[0571] The following abbreviations were used as noted: [0572] MeOH: methanol [0573] NaHCO.sub.3: sodium bicarbonate [0574] K.sub.2CO.sub.3: potassium carbonate [0575] MS: mass spectrometry [0576] DMSO: dimethyl sulfoxide [0577] TLC: thinlayer chromatography [0578] Et.sub.3N: triethylamine [0579] EtOAc: ethyl acetate [0580] DCM: dichloromethane [0581] NH.sub.4Cl: ammonium chloride [0582] THF: tetrahydrofuran [0583] Na.sub.2CO.sub.3: sodium carbonate [0584] EDCI: N-Ethyl-N′-(3-dimethylaminopropyl)carbodiimide hydrochloride [0585] DMAP: 4-dimethylamino pyridine [0586] HATU O-(7-Azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium-hexafluorphosphat [0587] DIPEA N,N-Diisopropylethylamine

CITATION LIST

Patent Literature

[0588] US2007238882A1

[0589] US2003105066A1

[0590] US2008221158A1

[0591] US2008027012A1

[0592] US6455576B1

[0593] US5677287A

[0594] EP1748994B1

[0595] WO2007025632A2

[0596] WO9530641A1

[0597] WO0002567A1

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