NO322424B1 - Derivatives of 4'-demycarosyl-8a-aza-8a-homothylosin - Google Patents

Description

The present invention relates to new compounds from the class of 17-membered azalides which have an antibacterial effect. More particularly, the invention relates to a compound characterized by the general formula I

where R represents CHO, CH(OCH3)2 or CH2N[CH2(C6H5)]2,

R1 represents C1-C3acyl or R<1>represents H when R<2>and R3 orR<4>and R<5>together are =0,

R<2>represents OR6 and R<6>represents H or C1-C3acyl,

R<3>represents H or R2 and R3 together represent =0,

R<4>represents OH,

R 5 represents H or R 4 and R 5 together represent =0.

Known technique

4'-Demycarosyl-8a-aza-8a-homotylosin, a new semisynthetic macrolide from the class of 17-membered azalides, was prepared by a double transformation of the C-9 ketone in the 16-membered antibiotic 4'-demycarosyl-tylosin (R. L. Hamill, Antibiotics and Chemotherapy 11, 328 (1961); A. Narandja et al, EP 0 287 082 Bl; N. Lopotar et al, EP 0 410 433 Bl). By reductive amination of the C-20 aldehyde group in the presence of formic acid (Wallach reaction, J. March: "Advanced Organic Chemistry", third ed. 6-15 p. 799 Wiley, New York, 1985) 4'- demycarosyl-20-deoxo-20-dibenzylamino-8a-aza-8a-homotylosin (N. Lopotar, HR patent application P940962A, 30.11.1994).

C1-C3 acyl esters of 4'-demycarosyl-8a-aza-8a-homotylosin and of 4'-demycarosyl-20-deoxo-20-dbenzylairiino-8a-aza-8a-homotylosin as well as 4".-deoxy-4"- oxo- and 3-deoxy-3-oxo-derivatives of 4'-demycarosyl-8a-aza-8a-homotylosin and of 4*-demycarosyl-20-deoxo-20-dibenzylamino-8a-aza-8a-homotylosin, Q -C3acyl esters thereof and a method for their production have not been described in the prior art so far.

Detailed description of the invention

Derivatives of 4'-demycarosyl-8a-aza-8a-homotylosin of formula I

where R represents CHO, CH(OCH3)2 or CH2N[CH2(C6H5)]2,

R<1> represents H or C1-C3 acyl, .

R represents OR and R represents H or C1-C3acyl,

R<3>represents H or R2 and R3 together represent =0,

R<4>represents OH,

R<5> represents H or R4 and R<5> together represent =0,

is produced by

4'-demycarosyl-8a-aza-8a-homotylosin 20-dimethylacetal of formula Ha and 4'-demycarosyl-20-deoxo-20-dibenzylamino-8a-aza-8a-homotylosin of formula Hb

Ea R = CH(OCH 3 ) 2

Hb R = CH 2 N[CH 2 (C 6 H 5 )] 2

can be submitted

A) an O-acylation with anhydrides of C1-C3 carboxylic acids, preferably with acetic anhydride in methylene chloride for 15 minutes to 1 hour at room temperature and the obtained compounds of formula I, where R represents CH(OCH3)2 or CH2N[CH2(C6H5)]2 , R<1>represents COCH3, R2 represents OR<6>, where R<6>represents H, R3 and R<5>are equal and represent H and R<4>represents OH,

may possibly be submitted

Al) O-acylation with anhydrides of C1-C3 carboxylic acids, preferably with acetic anhydride in methylene chloride in the presence of an organic base, preferably triethylamine and 4-dimethylaminopyridine as catalyst for 30 hours at room temperature, and the obtained compounds of formula I, where R represents CH (OCH3)2or CH2N[CH2(C6H5)]2, R<1>represents COCH3, R2 represents OR<6>, where R<6>represents COCH3, R3 and R<5>are equal and represent H and R<4 >represents OH,

may possibly be submitted

B) an oxidation reaction with N(3-dimethylaminopropyl)-N'ethylcarbodiimide hydrochloride in the presence of dimethyl sulfoxide and pyridine trifluoroacetate as a catalyst in an inert solvent, preferably methylene chloride, for 2 to 6 hours at a temperature from 10°C to room temperature, and the obtained compounds of formula I, where R represents

CH(OCH3)2or CH2N[CH2(C6H5)]2, R<1>represents COCH3, R<2>represents OR<6>, where R represents COCH3, R represents H and R and R together represent =0,

may possibly be submitted

C) methanolysis at room temperature for 2 days, and the obtained compounds of formula I, where R represents CH(OCH3)2 or CH2N[CH2(C6H5)]2, R<1> and R<3> are equal and represent H, R<2>represents OR<6>, where R<6>represents COCH3 and R<4>and R<5>together represent =0,

may possibly be submitted

Cl) alkaline methanolysis in a mixture of methanol and 25% ammonia (4:1) at a temperature from 5°C to room temperature for 20 to 60 hours to obtain compounds of formula I, where R represents CH(OCH3)2 or CH2N[ CH2(C6H5)]2, R<1>and R<3>are equal and represent H, R<2>represents OR<6>, where R<6>represents H and R<4>andR<5>together represent =0;

or the compound obtained according to process Cl

of formula I, where R represents CH(OCH3)2, R<1>and R<3>are equal and represent H, R<2>represents OR<6>, where R<6>represents H and R<4> and R<5> together represent =0,

may possibly be submitted

D) hydrolysis of the acetal in a mixture of acetonitrile and 0.1 N hydrochloric acid (1:1) for 2 hours at room temperature to obtain the compound of formula I, where R represents a CHO group, R<1> and R3 are equal and represents H, R2 represents OR<6>, where R<6>represents H and R<4>and R<5>together represent =0;

or compounds obtained according to process A

of formula I, where R represents CH(OCH3)2 or CH2N[CH2(C6H5)]2, R<1>represents COCH3, R<2>represents OR<6>, where R<6>represents H, R<3> and R<5>are equal and represent H and R<4>represent OH,

may possibly be subjected to oxidation in the same way as described in B,

and the obtained compounds of formula I, where R represents CH(OCH3)2 or CH2N[CH2(C6H5)]2, R<1>represents COCH3, R<2>and R<3> together represent =0, R<4> represents OH and R<5>represents H,

may optionally be subjected to methanolysis in the same way as described in C,

to obtain compounds of formula I, where R represents CH(OCH 3 ) 2 or CH 2 N[CH 2 (C 6 H s )] 2 , R and R are equal and represent H, R and R together represent =O and R<4> represents OH;

or the compound obtained according to process B

with formula I, where R represents a CH(OCH3)2 group, R<1>represents COCH3, R<2>and R3 together represent =0, R<4>represents OH and R<5>represents H,

may optionally be subjected to hydrolysis of the acetal in the same manner as described in D, and the obtained compound of formula I, where R represents a CHO group, R<1> represents COCH3, R2 and R<3> together represent =0, R <4>represents OH and R<5>represents H,

may optionally be subjected to methanolysis in the same way as described in C,

to obtain the compound of formula I, where R represents a CHO group, R<1> and R<5> are equal and represent H, R2 and R<3> together represent =0 and R4 represents OH;

or the compound obtained according to process A

of formula I, where R represents CH(OCH3)2, R<1>represents COCH3, R<2>represents OR<6>, where R6 represents H, R3 and R<5>are equal and represent H and R<4 >represents OH,

may optionally be subjected to hydrolysis of the acetal in the same manner as described in D, to obtain a compound of formula I where R represents CHO, R<1> represents COCH3, R2 represents OR<6>, where R6 represents H, R3 and R< 5> are equal and represent H and R 4 represents OH;

or compounds obtained according to process Al

of formula I, where R represents CH(OCH3)2 or CH2N[CH2(C6H5)]2, R<1>represents COCH3, R<2>represents OR<6>, where R<6>represents COCH3, R3 and R< 5>are equal and represent H and R<4>represents OH,

may optionally be subjected to methanolysis in the same way as described in C,

to obtain compounds of formula L where R represents CH(OCH3)2 or CH2N[CH2(C6H5)]2, R<1>, R3 and R<5> are equal and represent H, R<2> represents OR<6> , where R<6>represents COCH3 and R<4>represents OH;

or the compound obtained according to process Al

of formula I, where R represents CH(OCH3)2, R<1>represents COCH3, R<2>represents OR<6>, where R<6>represents COCH3, R<3>and R<5>are equal and represents H and R<4>represents OH,

may optionally be subjected to hydrolysis of the acetal in the same manner as described in D, and the obtained compound of formula I, where R represents CHO, R<1>represents COCH3, R<2>represents OR<6>, where R6 represents COCH3, R3 and R<5>are equal and represent H and R<4>represents OH,

may optionally be subjected to methanolysis in the same way as described in C,

to obtain the compound of formula I, where R represents CHO, R<1>, R<3> and R<5> are equal and represent H, R<2> represents OR<6>, where R<6> represents COCH3 and R<4> represents OH.

New compounds can be isolated by conventional processes by extraction from aqueous solutions of halogenated hydrocarbons such as methylene chloride or chloroform and by evaporation of the organic solvent to a dry residue. Optionally, the separation of the reaction products or the purification of the products for spectral analyzes is carried out by "flash" chromatography on a silica gel column (Merck & Co., Silicagel 60,230-400 mesh/ASTM) in a solvent system: CH2Cl2-CH3OH-conc. NH 4 OH (90:9:1.5, system A), CH 2 Cl 2 -CH 3 OH (90:9, system B) or CHCl 3 -CH 3 COCH 3 (7:3, system C).

The structure of the new compounds was confirmed by spectrometric methods and mass analysis.

The new compounds exhibit antibacterial activity and can also be used as intermediates for the production of new 17-membered azalide antibiotics.

The invention is illustrated in the following Examples.

Example 1

4'-Demycarosyl-2',4'-di-0-acetyl-8a-aza-8a-homotylosine 20-dimethyl acetal (1)

4'-Demycarosyl-8α-aza-8α-homotylosin-20-dimethyl acetal (5.0 g, 6.02 mmol) was dissolved in dry methylene chloride (50 mL), acetic anhydride (2.0 mL) was added and it was stirred for 15 minutes at room temperature. The reaction mixture was poured into a water/ice mixture (500 mL) and extracted twice with methylene chloride at pH 8.5. The combined organic extracts were washed with a saturated NaHCO 3 solution and water, dried (K 2 CO 3 ) and evaporated under reduced pressure to give a TLC homogeneous product (1) (5.38 g; 97.8<%>)<.>

TLC: R f (B) 0.44; R f (C) 0.22.

IRfKBOcm"<1>1749, 1657,1620,1544,1455,1375,1229,1170,1063.

1 H NMR (CDCl 3 ) 8 ppm 7.16 (H-11), 5.69 (H-10), 5.66 (H-13), 4.96 (8a-NH) exchangeable with D 2 O, 4.88 (H-2'), 4.76 (H-4'), 4.63 (H-20), 4.58 (H-l"), 4.33 (H-l*), 4.17 (H-8) , 3.61 (3"-OCH3), 3.47 (2"-OCH3), 3.56 (2x20-OCH3), 2.33 /3'-N(CH3)2/,

2.05 (COCH 3 ), 2.03 (COCH 3 ), 1.74 (H-22), 1.17 (H-21).

<13>C NMR (CDCl3) 8 ppm 179.1 (C-1), 169.8, 169.4 (2xCOCH3), 166.2 (9-CONH),

144.7 (C-11), 138.2 (C-13), 134.9 (C-12), 119.2 (C-10), 103.5 (C-20), 102.0 (C -1'), 100.9 (C-1"), 72.5 (C-4"), 71.4 (C-4'), 70.3 (C-2<*>), 65.6 (C -3), 61.5 (3"-OCH3), 59.4 (2"-OCH3), 50.4 (2x20-OCH3), 42.7 (C-8), 42.5 (C-4) , 41.0 /3'-N(CH3)2/, 40.5 (C-2), 34.3 (C-19), 21.8, 20.9 (2xCOCH3), 21.9 (C- 21), 12.6 (C-22), 8.3 (C-18).

FAB (MH<+>) 917.

Example 2

4'-Demycarosyl-2',4'-di-O-acetyl-20-deoxo-20-dibenzylamino-8a-aza-8a-homotylosin (2)

4'-Demycarosyl-20-deoxo-20-dibenzylamino-8a-aza-8a-homotylosin (2.8 g, 2.90 mmol) was dissolved in dry methylene chloride (30 mL), acetic anhydride (1.3 mL, 13, 76 mmol) was added and it was stirred for 15 minutes at room temperature. The reaction mixture was poured into a water/ice mixture (300 mL) and extracted twice with methylene chloride at pH 6.5. The combined organic extracts were washed with a saturated NaHCC«3 solution and

water, dried (K2CO3) and evaporated under reduced pressure to give a TLC homogeneous product (2) (3.02 g; 98.9%).

TLC: R f (B) 0.38; R f (C) 0.23.

IRtKB^cm-1 1749, 1651, 1633, 1548, 1454, 1374, 1231, 1169, 1059.

1 H NMR (CDCl 3 ) 8 ppm 7.25 ~ 7.41 (phenyl), 7.10 (H-1 1), 5.70 (H-13), 5.65 (H-10), 4.89

(8a-NH) replaceable by D2O, 4.84 (H-2'), 4.74 (H-4'), 4.60 (H-l"), 4.15 (H-l'), 3.62 (3"-OCH3), 3.61 (20-N-CH2-phenyl), 3.58 (20-CH2-phenyl), 3.51 (2"-OCH3), 2.32 /3'-N( CH 3 ) 2 /, 2.06 (COCH 3 ), 2.00 (COCH 3 ), 1.72 (H-22), 1.12 (H-21).

,<3>C NMR (CDCl3) 8 ppm 173.4 (C-1), 169.9, 169.5 (2xCOCH3), 166.1 (9-CONH),

144.8 (C-11), 137.9 (C-13), 135.2 (C-12), 119.3 (C-10), 102.3 (C-1'), 101.0 ( C-l"), 72.5 (C-4"), 71.4 (C-4'), 70.4 (C-2'), 66.0 (C-3), 61.5 (3"- OCH3), 59.5 (2"-OCH3), 52.2 (C-20), 42.9 (C-8), 42.4 (C-4), 41.0 /3'-N(CH3 )2/, 38.7 (C-2), 29.4 (C-19), 21.8 (C-21), 21.1, 21.0 (2xCOCH3), 12.7 (C-22) , 8.4 (C-18), 20-N(CH 2 C 6 H 5 ) 2 , 139.8, 129.1, 128.0, 126.6, 57.9.

FAB (MH<+>) 1052.

Example 3

4'-Demycarosyl-2',4',4"-tri-O-acetyl-8a-aza-8a-homothyosine-20-dimethyl acetal

(3)

Compound 1 (4.0 g, 4.37 mmol) was dissolved in dry methylene chloride (100 mL), triethylamine (7.0 mL), 4-dimethylaminopyridine (0.12 g) and acetic anhydride (0.42 mL, 4, 45 mmol) was added and then the reaction solution was allowed to stand for 26 hours at room temperature. Isolation of the product was carried out in the same manner as described in Example 1, which gave a TLC homogeneous product (3) (4.08 g; 97.7%).

TLC: R f (A) 0.65; R f (C) 0.54.

IR (KBr) cm"<1>1749,1655,1618,1546,1454,1374,1233,1171,1052.

1 H NMR (CDCl 3 ) 8 ppm 7.16 (H-1 1), 5.69 (H-10), 5.65 (H-13), 4.89 (8α-NH) exchangeable with D 2 O, 4.88 ( H-2'), 4.76 (H-4'), 4.64 (H-1"), 4.59 (H-20), 4.33 (H-1'), 4.18 (H-8 ), 3.52 (3"-OCH3), 3.46 (2"OCH3), 3.36 (20-OCH3), 3.35 (20-OCH3), 2.33 /3'-N(CH3) 2/, 2.12 (COCH3), 2.05 (COCH3), 2.03 (COCH3), 1.74 (H-22), 1.16

(H-21).

<13>C NMR (CDCl3) 8 ppm 173.1 (C-1), 170.1, 169.8, 169.4 (3xCOCH3), 166.1

(9-CONH), 144.7 (C-11), 138.0 (C-13), 134.9 (C-12), 119.2 (C-10), 103.7 (C-20), 102.1 (C-1'), 100.9 (C-1"), 74.5 (C-4"), 71.4 (C-4'), 70.3 (C-2')„ 65, 6 (C-3), 61.3 (3"-OCH3), 59.3 (2"-OCH3), 53.7 (20-OCH3), 50.6 (20-OCH3), 42.7 (C -8), 42.6 (C-4), 41.0/3'-N(CH3)2/, 40.5 (C-2), 34.5 (C-19), 21.9, ( C-21), 21.1,

21.0, 20.7 (3xCOCH3), 12.7 (C-22), 8.3 (C-18).

FAB (MH<+>) 959.

Example 4

4'-Demycarosyl-2',4',4"-tri-O-acetyl-20-deoxo-20-dibenzylamino-8a-aza-8a-homotylosin (4)

Compound 2 (2.8 g, 2.66 mmol) was dissolved in dry methylene chloride (60 mL), triethylamine (3.7 mL), 4-dimethylaminopyridine (0.07 g) and acetic anhydride (0.25 mL, 1, 64 mmol) was added and then the reaction solution was allowed to stand for 26 hours at room temperature. Isolation of the product was carried out in the same manner as described in Example 1, which gave a TLC homogeneous product (4) (2.7 g; 92.9%).

TLC: Rf (B) 0.55; R f (C) 0.47.

IR (KBr) cm"<1>1747,1651,1632,1538, 1453,1372,1233,1170,1051.

1 H NMR (CDCl 3 ) 8 ppm 7.22 ~ 7.41 (phenyl), 7.10 (H-1 1), 5.70 (H-13), 5.65 (H-10),

4.91 (8a-NH) exchangeable with D20, 4.86 (H-2'), 4.74 (H-4'), 4.66 (H-l"), 4.46 (H-4"), 4.15 (H-1'), 3.61 (2x20-N-CH2-phenyl), 3.53 (3"-OCH3), 3.50 (2"-OCH3), 2.32 /3'- N(CH3)2/, 2.12 (COCH3), 2.06 (COCH3), 2.00 (COCH3), 1.72 (H-22),

1.12 (H-21), 0.78 (H-18).

<13>C NMR (CDC13) 8 ppm 173.3 (C-1), 170.1, 169.9, 169.5 (3xCOCH3), 166.1

(9-CONH), 144.8 (C-11), 137.9 (C-13), 135.2 (C-12), 119.3 (C-10), 102.3 (C-1' ),

101.0 (C-1"), 74.6 (C-4"), 71.4 (C-4'), 70.4 (C-2<*>), 66.0 (C-3), 61 .5 (3"-OCH3), 59.3 (2"-OCH3), 52.2 (C-20), 42.9 (C-8), 42.4 (C-4), 41.0 / 3*-N(CH3)2/, 38.7 (C-2), 29.4 (C-19), 21.8 (C-21), 21.1,21.0,20.7 (3xCOCH3 ), 12.7 (C-22), 8.4 (C-18),

20-N(CH 2 C 6 H 5 ) 2.139.8.129.1.128.0, 126.6.57.9.

FAB (MH<+>) 1094.

Example 5

4'-Demycarosyl-2',4'-di-O-acetyl-4"-deoxy-4"-oxo-8a-a2a-8a-homotylosin-20-dimethyl acetal (5)

A solution of pyridine trifluoroacetate (1.0 g, 5.24 mmol) in methylene chloride (10 mL) was added dropwise at 15 °C to a solution of compound 1 (1.0 g, 1.09 mmol), l- (3-Dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.0 g, 5.22 mmol) and dimethyl sulfoxide (1.0 mL, 14.10 mmol) in methylene chloride (20 mL). The reaction mixture was stirred for 3 hours at room temperature, then poured into water (150 ml) and after separation of the organic layer, it was extracted twice more with methylene chloride. The combined organic extracts were washed with a saturated NaHCO 3 solution and water, dried (K 2 CO 3 ) and evaporated under reduced pressure to a dry residue. The crude product obtained (0.95 g) was purified by flash chromatography on a silica gel column using the solvent system B to give a TLC homogeneous product (5) (0.45 g).

TLC: Rf (B) 0.52.

IR (KBr) cm'1 1749, 1657, 1620, 1542, 1455, 1375, 1230, 1172, 1060.

1 H NMR (CDCl 3 ) 8 ppm 7.16 (H-11), 5.71 (H-10), 5.64 (H-13), 4.97 (8a-NH) exchangeable with D 2 O, 4.88 (H-2'), 4.76 (H-4'), 4.60 (H-20), 4.63 (H-l"), 4.33 (H-l'), 4.17 (H- 8), 3.98 (H-5"), 3.78 (H-3"), 3.58 (3"-OCH3), 3.52 (2"-OCH3), 3.36 (20-OCH3 ), 3.35 (20-OCH3), 3.30 (H-2"), 2.33 /3'-N(CH3)2/, 2.05 (COCH3), 2.03 (COCH3), 1 .76 (H-22), 1.34 (H-6"), 1.17 (H-21).

<13>C NMR (CDC13) 8 ppm 202.4 (C-4"), 173.1 (C-1), 169.9, 169.5 (2xCOCH3), 166.1 (9-CONH), 144.6 (C-1 1), 137.6 (C-13), 135.3 (C-12), 119.5 (C-10), 103.6 (C-20), 103.0 (C-1"), 102.1 (C-P), 85.3 (C-3"), 84.2 (C-2"), 73.3 (C-5"), 71.3 (C-4'), 70.3 (C- 2'), 65.6 (C-3), 60.2 (3"-OCH3), 59.1 (2"-OCH3), 53.7 (20-OCH3), 50.5

(20-OCH3), 42.7 (C-8), 42.6 (C-4), 41.0 /3'-N(CH3)2/, 40.7 (C-2) 34.4 ( C-19), 21.9 (C-21), 21.1,21.0 (2xCOCH3), 14.0 (C-6"), C-12.7 (C-22), 8.3 ( C-18).

FAB(MH<+>)915.

Example 6

4,-Demycarosyl-2',4'-di-O-acetyl-4"-deoxy-4?,-oxo-20-deoxo-20-dibenzylamino-8a-aza-8a-homotylosin (6)

A solution of pyridine trifluoroacetate (0.6 g, 3.11 mmol) in methylene chloride (6 mL) was added dropwise at 15 °C to a solution of compound 2 (0.6 g, 0.57 mmol), l-(3- dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (0.6 g, 3.14 mmol) and dimethyl sulfoxide (0.45 mL, 6.35 mmol) in methylene chloride (20 mL). The reaction mixture was stirred for 5 hours at room temperature. Isolation of the product was carried out in the same manner as described in Example 5. The obtained crude product (0.54 g) was purified by "flash" chromatography on a silica gel column using solvent system B, which gave a TLC homogeneous product (6) (0 .28 g).

TLC: R f (B) 0.48; R f (C) 0.33.

IR (KBr) cm-1 1747, 1651, 1633, 1548, 1454, 1372, 1231, 1058.

1 H NMR (CDCl 3 ) 8 ppm 7.25 ~ 7.41 (phenyl), 7.12 (H-1 1), 5.70 (H-13), 5.65 (H-10), 4.94

(8a-NH) replaceable by D20,4,82 (H-2*), 4.74 (H-4'), 4.65 <H-1"), 4.15 (H-1'), 3 .98 (H-5"), 3.78 (H-3"), 3.62 (20-N-CH2-phenyl), 3.58 (20-CH2-phenyl), 3.55 (3"- OCH3), 3.49 (2"-OCH3), 2.32 /3'-N(CH3)2/, 2.06 (COCH3), 2.00 (COCH3), 1.74

(H-22), 1.36 (H-6"), 1.12 (H-21).

<13>C NMR (CDC13) 8 ppm 202.4 (C-4"). 173.4 (C-1), 169.8, 169.3 (2xCOCH3), 166.1 (9-CONH), 144.6 (C-1 1), 137.0 (C-13), 135.6 (C-12), 119.6 (C-10), 103.0 (C-l"), 102.2 (C-r), 85.3 (C -3"), 84.8 (C-2"), 73.3 (C-5"), 71.4 (C-4"), 70.4 (C-2'), 65.9 (C -3), 60.3 (3"-OCH3), 59.1 (2"-OCH3), 52.2 (C-20), 42.9 (C-8), 42.4 (C-4) , 40.9 /3'-N(CH3)2/, 38.7 (C-2), 29.4 (C-19), 21.8 (C-21), 21.1, 21.0 ( 2xCOCH3), 14.0 (C-6"), 12.8 (C-22), 8.4 (C-18),

20-N(CH2C6H5)2139.6.129.9.128.0.126.6.57.8.

FAB (MH<+>) 1050.

Example 7

4'-Demycarosyl-2',4',4"-tri-O-acetyl-3-deoxy-3-oxo-8a-aza-8a-homotylosin-20-dimethyl acetal (7)

A solution of pyridine trifluoroacetate (3.0 g, 15.72 mmol) in methylene chloride (30 mL) was added dropwise at 15 °C to a solution of compound 3 (2.0 g, 2.09 mmol), l-( 3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (3.0 g, 15.66 mmol) and dimethyl sulfoxide (2.9 mL, 40.89 mmol) in methylene chloride (50 mL).The reaction mixture was stirred for 3 hours at room temperature. Isolation of the product was carried out in the same manner as described in Example 5. The crude product obtained (1.95 g) was purified by "flash" chromatography on a silica gel column using the solvent system C, which gave a TLC homogeneous product (7) (1 .3 g).

TLC: R f (C) 0.58.

IR (KBr) cm 1 1749, 1655, 1618, 1546, 1454, 1374, 1233, 1171, 1052.

1 H NMR (CDCl 3 ) 8 ppm 6.90 (H-1 1 ), 5.76 (H-10 ), 5.43 (H-13 ), 4.96 (8a-NH) exchangeable with D 2 O , 4.89 ( H-2'), 4.79 (H-4'), 4.66 (H-l"), 4.40 (H-l<*>), 4.18 (H-8), 3.55, 3.32 (H-2), 3.52 (3"-OCH3), 3.49 (2"-OCH3), 3.30 (20-OCH3), 3.29 (20-OCH3), 2.34/3' -N(CH3)2/, 2.12 (COCH3), 2.06 (COCH3), 2.03 (COCH3), 1.75 (H-22), 1.10 (H-21), 1.07 (H-18).

<13>C NMR (CDCl3) 8 ppm 205.6 (C-3), 172.9 (C-1), 170.1,169.8,169.4 (3xCOCH3),

166.1 (9-CONH), 144.1 (C-11), 138.0 (C-13), 134.9 (C-12), 119.6 (C-10), 103.7 (C -20), 102.1 (C-l<*>), 100.9 (C-l"), 74.5 (C-4"), 71.4 (C-4<*>), 70.3 (C- 2'), 61.3 (3"-OCH3), 59.3 (2"-OCH3), 53.7 (20-OCH3), 50.6 (20-OCH3), 46.5 (C-2) , 44.2.

(C-4), 42.0 (C-8), 41.0 /3'-N(CH3)2/, 34.5 (C-19), 21.9, (C-21), 21, 1.21.0.20.7 (3xCOCH3), 17.6 (C-18), 12.7 (C-22).

FAB (MH<+>) 957.

Example 8

4'-dem<y>karos<y>l-2',4',4"-tri-O-acet<y>l-3-deox<y>-3-oxo-20-deoxo-20-dibenz <y>laniino-8a-aza-8a-homotylosin (8)

A solution of pyridine trifluoroacetate (2.0 g, 10.36 mmol) in methylene chloride (10 mL) was added dropwise at 15 °C to a solution of compound 4 (1.0 g, 1.09 mmol), l- (3-Dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (2.04 g, 10.44 mmol) and dimethyl sulfoxide (1.6 mL, 22.56 mmol) in methylene chloride (20 mL). The reaction mixture was stirred for 6 hours at room temperature. The isolation of the product was carried out in the same manner as described in Example 5. The obtained crude product (0.96 g) was purified by "flash" chromatography on a silica gel column using the solvent system B, which gave a TLC homogeneous product (8) (0 .62 g).

TLC: Rf (B) 0.60.

IR (KBr) cm 1 1748, 1633, 1538, 1454, 1373, 1231, 1052.

1 H NMR (CDCl 3 ) 5 ppm 7.22 - 7.40 (phenyl), 6.89 (H-1 1), 5.66 (H-10), 5.49 (H-13),

4.96 (8a-NH) exchangeable with D20, 4.81 (H-2'), 4.74 (H-4'), 4.66 (H-l"), 4.42 (H-4"), 4.15 (H-1'), 4.12 (H-8), 3.78, 3.38 (H-2), 3.51 (2x20-N-CH2-phenyl, 3"-OCH3), 3.48 (2"-OCH3), 2.32 /3'-N(CH3)2/, 2.22 (H-4), 2.09 (COCH3), 2.06 (COCH3), 2.00 (COCH 3 ), 1.72 (H-22), 1.10 (H-21), 1.08 (H-18).

<13>C NMR (CDCl3) 5 ppm 206.7 (C-3), 172.7 (C-1), 170.1,169.9,169.5 (3xCOCH3),

166.1 (9-CONH), 144.0 (C-II), . 136.5 (C-12), 135.0 (C-13), 119.9 (C-10), 102.7 (C-l'), 100.9 (C-l"), 74.6 (C -4"), 71.3 (C-4<*>), 70.3 (C-2'), 61.3 (3"-OCH3), 59.3 (2"-OCH3), 51.7 (C-20), 47.7 (C-2), 44.5 (C-4)), 42.0 (C-8), 41.0/3'-N(CH3)2/, 28, 6 (C-19), 22.0 (C-21), 21.0,20.7 (3xCOCH3), 17.8 (C-18), 13.1 (C-22),

20-N(CH 2 C 6 H 2 ), 140.1, 128.9, 128.0, 126.4, 57.9.

FAB (MH<+>) 1092.

Example 9

4,-Demycarosyl-4"-deoxy-4"-oxo-8a-aza-8a-homotylosm 20-dimethyl acetal (9)

Compound 5 (0.65 g, 0.71 mmol) was dissolved in methanol (20 mL) and allowed to stand at room temperature for 48 h. To the reaction solution was added a saturated NaHCO 3 solution and it was extracted twice with chloroform. The combined organic extracts were dried (K2CO3) and evaporated under reduced pressure to a dry residue. The crude product obtained (0.45 g) was purified by flash chromatography on a silica gel column using solvent system A, giving a TLC homogeneous product (9) (0.20 g).

TLC: Rf (A) 0.27.

JR (KBr) cm"1 1749,1657,1620,1542,1455,1375,1230,1172,1060.

1 H NMR (CDCl 3 ) 8 ppm 7.16 (H-1 1 ), 5.72 (H-10), 5.67 (H-13), 4.99 (8α-NH) exchangeable with D 2 O, 4.60 ( H-20), 4.63 (H-l"), 4.33 (H-l'), 4.17 (H-8), 3.98 (H-5"), 3.78

(H-3"), 3.58 (3"-OCH3), 3.52 (2"-OCH3), 3.46 (H-2'), 3.36, 3.35 (2x20-OCH3), 3.30 (H-2"), 3.06 (H-4'), 2.33 /3'-N(CH3)2/, 1.76 (H-22), 1.34 (H-6 "), 1.17 (H-21).

13 C NMR (CDCl 3 ) 8 ppm 202.4 (C-4"), 173.1 (C-1), 166.1 (9-CONH), 144.6 (C-1 1),

137.6 (C-13), 135.3 (C-12), 119.5 (C-10), 103.6 (C-20), 103.0 (C-l"), 102.1 (C- 1'), 85.3 (C-3"), 84.2 (C-2"), 73.3 (C-5"), 65.6 (C-3), 60.2 (3"- OCH3), 59.1 (2"-OCH3), 53.7 (20-OCH3), 50.5 (20-OCH3), 42.7 (C-8), 42.6 (C-4), 41 ,0 /3'-N(CH3)2/, 40.7 (C-2), 34.4 (C-19), 21.9 (C-21), 14.0 (C-6"), 12.7 (C-22), 8.3 (C-18).

FAB(MH<+>)831.

Example 10

4'-Demycarosyl-4"-deoxy-4"-oxo-20-deoxo-20-dibenzylamino-8a-aza-8a-homotylosin (10)

Compound 6 (0.30 g, 0.73 mmol) was dissolved in methanol (20 mL) and allowed to stand at room temperature for 30 h. After addition of water (50 ml), the product was isolated by gradient extraction with chloroform at pH 4.5 and 7.5. The combined chloroform extracts at pH 7.5 were dried (K 2 CO 3 ) and evaporated under reduced pressure and the product obtained (0.17 g) was purified by flash chromatography on a silica gel column using solvent system A, giving a TLC homogeneous product (10)

(0.08g).

TLC: Rf (A) 0.49.

IR (KBr) cm"1 1715,1655,1619,1542,1454,1377,1168,1082.

"H NMR (CDCl 3 ) 8 ppm 7.25 ~ 7.41 (phenyl), 7.12 (H-l 1), 5.70 (H-13), 5.65 (H-10), 4.94

(8a-NH) replaceable by D2O, 4.84 (H-2'), 4.74 (H-4'), 4.60 (H-l"), 4.15 (H-l'), 3.98 (H-5"), 3.78 (H-3"), 3.62 (3"-OCH3), 3.61 (20-N-CH2-phenyl), 3.58 (20-CH2-

phenyl), 3.51 (2"-OCH3), 3.46 (H-2'), 3.01 (H-4'), 2.32 /3'-N(CH3)2/, 1.72 (H-22),

1.12 (H-21).

<13>C NMR (CDCl3) 8 ppm 202.4 (C-4"), 173.4 (C-1), 166.1 (9-CONH), 144.7 (C-11),

137.1 (C-13), 135.6 (C-12), 119.7 (C-10), 104.2 (C-l'), 103.0 (C-l"), 85.4 (C -3"), 84.9 (C-2"), 73.3 (C-5"), 66.4 (C-3), 59.8 (3"-OCH3), 58.6 (2" -OCH3), 52.2 (C-20), 43.3 (C-8), 42.3 (C-4), 41.5 /3'-N(CH3)2/, 38.7 (C -2), 29.4 (C-19), 22.0 (C-21), 14.1 (C-6"), 12.8 (C-22), 9.1 (C-18),

20-N(CH 2 C 6 H 5 ) 2 139.8, 129.1, 128.0, 126.6, 58.0.

FAB (MH<+>) 967.

Example 11

4'-Demycarosyl-4"-0-acetyl-3-deoxy-3-oxo-8a-aza-8a-homotylosln 20-dimethyl acetal (11)

Compound 7 (0.70 g, 0.73 mmol) was dissolved in methanol (50 mL) and allowed to stand at room temperature for 24 h. The isolation of the product was carried out in the same manner as described in Example 9 and the obtained crude product (0.62 g) was purified by "flash" chromatography on a silica gel column using solvent system A, which gave a TLC homogeneous product (11) (0 .40 g).

TLC: Rf (A) 0.44.

IR (KBr) cm"<1>1749,1657,1620,1544,1455,1375,1229,1170,1063.

1 H NMR (CDCl 3 ) 8 ppm 6.87 (H-1 1), 5.77 (H-10), 5.44 (H-13), 5.18 (8α-NH) exchangeable with D 2 O, 4.88 ( H-2'), 4.64 (H-l"), 4.44 (H-4"), 4.30 (H-l'), 4.17 (H-8), 3.93 (H-5 "), 3.89 (H-3"), 3.53 (3"-OCH3), 3.50, 3.26 (H-2), 3.48 (2"-OCH3), 3.30 ( 20-OCH3), 3.29 (20-OCH3), 2.53 /3'-N(CH3)2/, 2.12 (COCH3), 1.75 (H-22), 1.25

(H-18).

<13>C NMR (CDCl3) 8 ppm 205.4 (C-3), 172.9 (C-1), 170.1 (COCH3), 167.4 (9-CONH),

143.4 (C-11), 136.2 (C-12), 134.6 (C-13), 120.7 (C-10), 104.2 (C-1'), 103.9 ( C-20), 100.8 (C-l"), 74.5 (C-4"), 70.9 (C-2'), 70.5 (C-2'), 61.3 (3"- OCH3), 59.0 (2"-OCH3), 52.6 (20-OCH3), 52.1 (20-OCH3), 45.9 (C-2), 44.4 (C-4), 42 .5 (C-8), 41.4 /3'-N(CH3)2/, 33.8 (C-19), 22.0 (C-21), 20.7 (COCH3), 17.5 (C-18), 12.9 (C-22).

FAB (MH<+>) 873.

Example 12

4'-Demycarosyl-4"-O-acetyl-3-deoxy-3-oxo-20-deoxo-20-dibenzylamino-8a-aza-8a-homotylosin (12)

Compound 8 (1.20 g, 10.99 mmol) was dissolved in methanol (100 mL) and allowed to stand at room temperature for 24 hours. To the reaction solution was added water (100 ml) and it was extracted with methylene chloride at pH 6.5. The combined organic extracts were dried (K2CO3) and evaporated under reduced pressure and the crude product obtained (1.0 g) was purified by flash chromatography on a silica gel column using solvent system A to give a TLC homogeneous product (12) ( 0.52g).

TLC: R f (A) 0.65.

IR (KBr) cm"1 1745,1650,1622, 1537,1454,1373,1233,1166,1058.

1 H NMR (CDCl 3 ) 8 ppm 7.25 ~ 7.41 (phenyl), 6.90 (H-11), 5.67 (H-10), 5.52 (H-13), 4.98

(8a-NH) exchangeable with D20, 4.67 (H-l"), 4.45 (H-4"), 4.17 (H-l'), 4.02 (H-8), 3.61 ( 20-CH2-phenyl), 3.53 (3"-OCH3), 3.52 (20-CH2-phenyl), 3.50 (2"-OCH3), 3.76, 3.32 (H-2) , 2.52 /3'-N(CH 3 ) 2 /, 2.12 (COCH 3 ), 1.73 (H-22), 1.21 (H-18), 1.08 (H-21).

,<3>C NMR (CDCl 3 ) 8 ppm 205.3 (C-3), 172.5 (C-1), 170.1 (COCH 3 ), 167.2 (9-CONH),

143.9 (C-11), 135.9 (C-12), 135.4 (C-13), 120.0 (C-10), 103.9 (C-1'), 100.9 ( C-l"), 74.6 (C-4"), 70.7 (C-4'), 70.4 (C-2'), 61.3 (3"-OCH3), 59.3 (2" -OCH3), 51.6 (C-20), 46.1 (C-2), 44.5 (C-4), 43.3 (C-8), 41.5 /3'-N(CH3 )2/, 28.8 (C-19), 22.0 (C-21), 20.7 (COCH3), 17.8 (C-18), 12.9 (C-22), 20-N (CH 2 C 6 H) 2139.9, 128.8, 128.0, 126.5, 58.0.

FAB (MH<+>) 1008.

Example 13

4'-Demycarosyl-4"-O-acetyl-8a-aza-8a-homotylosin-20-dimethyl acetal (13)

Compound 3 (0.5 g, 0.52 mmol) was dissolved in methanol (20 mL) and allowed to stand at room temperature for 24 h. The isolation of the product was carried out in the same manner as described in Example 9 and the obtained crude product (0.43 g) was purified by "flash" chromatography on a silica gel column using the solvent system A, which gave a TLC homogeneous product (13) (0 .32 g).

TLC: Rf (A) 0.32.

IR (KBr) cm"<1>1739, 1656,1616,1541,1455,1376,1237,1170,1062.

1 H NMR (CDCl 3 ) 8 ppm 7.15 (H-1 1 ), 5.71 (H-10), 5.66 (H-13), 4.97 (8a-NH) exchangeable with D 2 O, 4.64 ( H-l"), 4.62 (H-20), 4.44 (H-4"), 4.24 (H-l'), 4.18 (H-8), 3.53 (3"-OCH3 ), 3.47 (2"-OCH3), 3.37 (20-OCH3), 3.36 (20-OCH3), 2.50 /3'-N(CH3)2/, 2.12 (COCH3) , 1.75 (H-22), 1.17 (H-21).

FAB (MH<+>) 875.

Example 14

4'-Demycarosyl-4"-O-acetyl-20-deoxo-20-dibenzylamino-8a-aza-8a-homotylosin

(14)

Compound 4 (0.75 g, 0.69 mmol) was dissolved in methanol (20 mL) and allowed to stand at room temperature for 24 h. The isolation of the product was carried out in the same manner as described in Example 12 and the obtained crude product (0.66 g) was purified by "flash" chromatography on a silica gel column using solvent system A, which gave a TLC homogeneous product (14) (0 .45 g).

TLC: Rf (A) 0.50.

IR (KBr) cm1 1740, 1657,1621,1538,1454, 1373,1236,1169, 1054.

1 H NMR (CDCl 3 ) 8 ppm 7.25 ~ 7.41 (phenyl), 7.10 (H-1 1), 5.69 (H-13), 5.65 (H-10), 4.96

(8a-NH) exchangeable with D20, 4.66 (H-l"), 4.45 (H-4"), 4.14 (H-8), 4.07 (H-l'), 3.59 ( 20-N-CH2-phenyl), 3.56 (20-CH2-phenyl), 3.53 (3"-OCH3), 3.50 (2"-OCH3),

2.49 /3'-N(CH 3 ) 2 /, 2.12 (COCH 3 ), 1.73 (H-22), 1.11 (H-21), 0.94 (H-18).

FAB (MH<+>) 1010.

Example 15

4% demycarosyl-3-deoxy-3-oxo-8a-aza-8a-homotylosin-20-dimethylacetaI (15)

Compound 11 (0.40 g, 0.46 mmol) was dissolved in a methanol/conc. NH4OH mixture (4:1.50 ml) and allowed to stand for 60 hours at a temperature of 5°C. The reaction solution was evaporated to an oily residue and then a product was isolated in the same manner as described in Example 9. The crude product obtained (0.25 g) was purified by flash chromatography on a silica gel column using solvent system A, which gave a TLC homogeneous product (15) (0.15 g).

TLC: Rf (A) 0.39.

IR (KBr) cm-1 1739, 1714, 1650, 1620, 1544, 1455, 1375, 1170, 1063.

1H NMR (CDCl3) 8ppm 6.87 (H-11), 5.77 (H-10), 5.44 (H-13), 5.18 (8a-NH) exchangeable with D2O, 4.60 (H -20), 4.64 (H-l"), 4.33 (H-l'), 4.17 (H-8), 3.93 (H-5"), 3.89 (H-3") , 3.53 (3"-OCH3), 3.50, 3.26 (H-2), 3.48 (2"-OCH3), 3.30 (20-OCH3), 3.29

(20-OCH 3 ), 2.33 /3'-N(CH 3 ) 2 /, 1.75 (H-22), 1.25 (H-18).

FAB (MH+) 831.

Example 16

4'-Demycarosyl-3-deoxy-3-oxo-20-deoxo-20-dibenzylamino-8a-aza-8a-homotylosin (16)

Compound 12 (0.78 g, 0.77 mmol) was dissolved in a methanol/conc. NH4OH mixture (4:1.50 ml) and allowed to stand for 24 hours at room temperature. To the reaction solution was added water (80 ml) and it was extracted twice with methylene chloride at pH 7.5. The combined organic extracts were dried (K 2 CO 3 ) and evaporated under reduced pressure and the product obtained (0.66 g) was purified by flash chromatography on a silica gel column using solvent system A to give a TLC homogeneous product (16)

(0.32g).

TLC: Rf (A) 0.55.

IR (KBr) cm-1 1739, 1714, 1650, 1622, 1538, 1454, 1376, 1167, 1082.

1 H NMR (CDCl 3 ) 8 ppm 7.25 ~ 7.41 (phenyl), 6.90 (H-1 1), 5.66 (H-13), 5.53 (H-10), 5.28

(8a-NH) replaceable by D20,4,61 (H-l"), 4.16 (H-l'), 4.03 (H-8), 3.62 (20-N-CH2-phenyl), 3 .61 (20-CH2-phenyl, 3"-OCH3), 3.51 (2"-OCH3), 3.78, 3.38 (H-2),

2.5 /3'-N(CH 3 ) 2 /, 2.38 (H-4), 1.72 (H-22), 1.21 (H-18), 1.08 (H-21).

,<3>C NMR (CDCl 3 ) 8 ppm 205.3 (C-3), 172.5 (C-1), 167.2 (9-CONH), 143.9 (C-11),

135.9 (C-12), 135.6 (C^13), 120.0 (C-10), 103.9 (C-1'), 101.0 (C-1'), 72.5 (C-4"), 70.7 (C-4'), 70.4 (C-2'), 61.5 (3"-OCH3), 59.5 (2"-OCH3), 51.7 (C-20), 46.1 (C-2), 44.5 (C-4), 43.3 (C-8), 41.5 /3'-N(CH3)2/, 28.8 (C-19), 22.0 (C-21), 17.8 (C-18), 12.9 (C-22),

20-N(CH 2 C 6 H) 2140.0, 128.8, 128.0, 126.5, 58.0.

FAB (MH<+>) 967.

Example 17

4'-Demycarosyl-3-deoxy-3-oxo-8a-aza-8a-homotylosin (17)

Compound 15 (0.5 g, 0.60 mmol) was dissolved in an acetonitrile/0.1 N HCl mixture (1:1, 35 mL) and stirred for 2 h at room temperature. To the reaction solution was added a saturated NaHCO 3 solution and it was extracted twice with methylene chloride. The combined organic extracts were dried (K 2 CO 3 ) and evaporated under reduced pressure and the product obtained (0.42 g) was purified by flash chromatography on a silica gel column using solvent system A to give a TLC homogeneous product (17)

(0.25g).

TLC: Rf (A) 0.35.

IR (KBr) cm 1 1739, 1719, 1657, 1620, 1545, 1455, 1376, 1169, 1082.

1 H NMR (CDCl 3 ) 8 ppm 9.78 (H-20), 7.19 (H-1 1), 5.72 (H-10), 5.70 (H-13), 5.06

(8a-NH) exchangeable with D20,4.58 (H-l"), 4.18 (H-l'), 4.23 (H-8), 3.68, 3.32 (H-2), 3 .62 (3"-OCH3), 3.49 (2"-OCH3), 2.49 /3'-N(CH3)2/, 1.75 (H-22), 1.25 (H-18) , 1.18 (H-21).

<13>C NMR (CDCl3) 8 ppm 205.3 (C-3), 203.8 (C-20), 173.5 (C-1), 166.9 (9-CONH),

145.1 (C-11), 138.2 (C-13), 135.1 (C-12), 129.3 (C-10), 103.7 (C-1'), 101.1 ( C-1'), 72.8 (C-4"), 71.0 (C-4'), 70.4 (C-2'), 61.5 (3"-OCH3), 59.5 ( 2"-OCH3),

46.6 (C-19), 46.1 (C-2), 44.5 (C-4), 43.3 (C-8), 41.5 /3'-N(CH3)2/, 22.4 (C-21),

17.8 (C-18), 12.9 (C-22).

FAB (MH<+>) 785.

Example 18

4,-Demycarosyl-2,,4,-di-O-acetyl-8a-aza-8a-homotylosin (18)

Compound 1 (0.5 g, 0.55 mmol) was dissolved in an acetonitrile/0.1 N HCl mixture (1:1.35 mL) and stirred for 2 h at room temperature. The isolation of the product was carried out in the same way as described in Example 17, which gave a TLC homogeneous product (18) (0.34 g).

TLC: Rf (B) 0.35.

IR (KBr) cm 1 1749, 1657, 1620, 1548, 1455, 1375, 1231, 1170, 1059.

1 H NMR (CDCl 3 ) 8 ppm 9.75 (H-20), 7.21 (H-1 1), 5.72 (H-10), 5.71 (H-13), 5.08

(8a-NH) replaceable by D20, 4.89 (H-2<*>), 4.74 (H-4'), 4.58 (H-l"), 4.26 (H-l<*>), 3 .61 (3"-OCH3), 3.49 (2"-OCH3), 2.33 /3'-N(CH3)2/, 2.05 (COCH3), 2.03 (COCH3), 1.74 (H-22), 1.18 (H-21).

<13>C NMR (CDC13) 8 ppm 203.6 (C-20), 173.3 (C-1), 169.9, 169.5 (2xCOCH3), 166.5 (9-CONH), 145.2 (C-1 1 ), 138.3 (C-13), 135.0 (C-12), 119.0 (C-10), 101.6 (C-l<*>), 100.9 (C-l"), 72.5 (C-4"), 70.6 (C-4'), 70.3 (C-2<*>), 65.6 (C-3), 61.5 (3"-OCH3), 59, 5 (2"-OCH3), 46.3 (C-19), 42.5 (C-8), 41.0 /3'-N(CH3)2/, 38.5 (C-2), 21 .6 (C-21), 21.1, 21.0 (2xCOCH 3 ), 12.7 (C-22), 8.1 (C-18).

FAB (MH<+>) 871.

Example 19

4'-Demycarosyl-2',4',4"-tri-O-acetyl-8a-aza-8a-homotylosin (19)

Compound 3 (0.5 g, 0.52 mmol) was dissolved in an acetonitrile/0.1 N HCl mixture (1:1.35 mL) and stirred for 2 h at room temperature. The isolation of the product was carried out in the same way as described in Example 17, which gave a TLC homogeneous product (19) (0.47 g).

TLC: R f (B) 0.60; Rf (C) 0.50.

IR (KBr) cm-<1>1748,1659,1621,1538, 1455,1373, 1232,1171,1052.

1 H NMR (CDCl 3 ) 8 ppm 9.74 (H-20), 7.16 (H-1 1), 5.69 (H-10), 5.65 (H-13), 4.89

(8a-NH) replaceable by D2O, 4.88 (H-2'), 4.76 (H-4<*>), 4.64 (H-l"), 4.44 (H-4"), 4 .33 (H-1'), 4.18 (H-8), 3.52 (3"-OCH3), 3.46 (2"-OCH3), 2.33 /3'-N(CH3)2 /,

2.12 (COCH 3 ), 2.05 (COCH 3 ), 2.03 (COCH 3 ), 1.74 (H-22), 1.16 (H-21).

<13>C NMR (CDCl3) Spppm 203.6 (C-20), 173.1 (C-1), 170.1, 169.8, 169.4 (3xCOCH3),

166.1 (9-CONH), 144.7 (C-11), 138.0 (C-13), 134.9 (C-12), 119.2 (C-10), 103.7 (C -20), 102.1 (C-1'), 100.9 (C-1"), 74.5 (C-4"), 71.4 (C-4'), 70.3 (C-2< »>), 65.6 (C-3), 61.3 (3"-OCH3), 59.3 (2"-OCH3), 46.3 (C-19), 42.7 (C-8) , 42.6 (C-4), 41.0

/3'-N(CH3)2/, 40.5 (C-2), 34.5 (C-19), 21.9 (C-21), 21.1,21.0,20.7 ( 3xCOCH3),

12.7 (C-22), 8.3 (C-18).

FAB(MH<+>)913.

Example 20

4'-Demycarosyl-2',4'-di-O-acetyl-4"-deoxy-4"-oxo-8a-aza-8a-homotylosin (20)

Compound 5 (0.7 g, 0.77 mmol) was dissolved in an acetonitrile/0.1 N HCl mixture (1:1.50 mL) and stirred for 1 hour at room temperature. The isolation of the product was carried out in the same manner as described in Example 17, which gave a TLC homogeneous product (20) (0.36 g).

TLC: Rf (B) 0.48.

IR (KBr) cm -1 1749, 1656, 1619, 1543, 1458, 1375, 1230, 1172, 1058.

1 H NMR (CDCl 3 ) 8 ppm 9.75 (H-20), 7.21 (H-1 1), 5.72 (H-10), 5.70 (H-13), 5.08

(8a-NH) replaceable by D2O, 4.88 (H-2'), 4.74 (H-4'), 4.58 (H-1"), 4.30 (H-1'), 4.17 (H-8), 3.98 (H-5"), 3.78 (H-3"), 3.58 (3"-OCH3), 3.48 (2"-OCH3), 3.30 ( H-2"), 2.33 /3'-N(CH3)2/, 2.05 (COCH3), 2.03 (COCH3), 1.76 (H-22), 1.34 (H-6 "),

1.17 (H-21).

<13>C NMR (CDCl3) 8 ppm 203.0 (C-20), 202.4 (C-4"), 173.1 (C-1), 169.9, 169.5

(2xCOCH3), 166.5 (9-CONH), 145.0 (C-11), 138.1 (C-13), 135.1 (C-12), 119.0 (C-10), 102 .1 (C-l"), 100.9 (C-l'), 85.3 (C-3"), 84.2 (C-2"), 73.3 (C-5"), 71.3

(C-4<*>), 70.3 (C-2<*>), 65.6 (C-3), 61.5 (3"-OCH3), 59.4 (2**-OCH3) , 46.3 (C-19), 42.5 (C-8), 41.0/3'-N(CH3)2/, 38.5 (C-2), 21.9 (C-21) , 21.1, 21.0 (2xCOCH 3 ), 14.0 (C-6**), 12.7 (C-22), 8.3 (C-1).

FAB(MH<+>)869.

Example 21

4'-Demycarosyl-4'*-O-acetyl-8a-aza-8a-homotylosin (21)

Compound 19 (0.30 g, 0.33 mmol) was dissolved in methanol (20 mL) and allowed to stand for 24 h at room temperature. The isolation of the product was carried out in the same way as

described in Example 9 and the obtained crude product (0.25 g) was purified by "flash" chromatography on a silica gel column using solvent system A, which gave a TLC homogeneous product (21) (0.19 g).

TLC: Rf (A) 0.28.

IR (KBr) cm 1 1749, 1657, 1620, 1544, 1455, 1375, 1229, 1170, 1063.

H NMR (CDCl 3 ) 8 ppm 9.78 (H-20), 7.20 (H-1 1), 5.72 (H-10), 5.70 (H-13), 5.12

(8a-NH) exchangeable with D2O, 4.88 (H-2'), 4.64 (H-l"), 4.44 (H-4"), 4.18 (H-1'), 4.12 (H-8), 3.93 (H-5"), 3.89 (H-3"), 3.53 (3"-OCH3), 3.48 (2"-OCH3), 2.49 / 3'-N(CH 3 ) 2 /, 2.12 (COCH 3 ), 1.75 (H-22).

FAB (MH<+>) 829.

Example 22

4*-demycarosyl-4"-deoxy-4"*oxo-8a-aza-8a-honiotylosin (22)

Compound 20 (0.23 g, 0.27 mmol) was dissolved in methanol (20 mL) and allowed to stand for 24 h at room temperature. The isolation of the product was carried out in the same way as

described in Example 9 and the crude product obtained (0.14 g) was purified by "flash" chromatography on a silica gel column using solvent system A to give a TLC homogeneous product (22) (0.095 g).

TLC: Rf (A) 0.30.

IR (KBr) cm-<1>1717,1655,1625,1542,1454,1378,1170,1062.

<]>H NMR (CDCl 3 ) 5 ppm 9.76 (H-20), 7.20 (H-1 1), 5.72 (H-10), 5.70 (H-13), 5.12

(8a-NH) exchangeable with D2O, 4.64 (H-l"), 4.33 (H-l'), 4.18 (H-8), 3.98 (H-5"), 3.78 ( H-3"), 3.58 (3"-OCH3), 3.46 (2"-OCH3), 3.30 (H-2"), 3.06 (H-4'), 2.33 / 3'-N(CH 3 ) 2 /, 1.74 (H-22), 1.34 (H-6"), 1.16 (H-21).

<13>C NMR (CDCl3) 8 ppm 203.7 (C-20), 202.5 (C-4"), 173.4 (C-1), 166.6 (9-CONH),

144.9 (C-11), 137.6 (C-13), 135.4 (C-12), 119.4 (C-10), 102.1 (C-1'), 100.9 ( C-1"), 71.4 (C-4'), 70.3 (C-2'), 66.3 (C-3), 61.5 (3"-OCH3), 59.7 (2"- OCH3), 46.2 (C-19), 42.7 (C-8), 42.1 (C-4), 41.5 /3'-N(CH3)2/, 39.8 (C- 2), 21.7 (C-21),

14.0 (C-6"), 12.7 (C-22), 8.7 (C-18).

FAB (MH<+>) 785.

Claims (20)

<I>1., Compound characterized by the general formula I where R represents CHO, CH(OCH3)2 or CH2N[CH2(C6H5)]2, R<1>represents C1-C3 acyl or R<1>represents H when R<2>and R3 or R<4>and R< 5>together is =0, R2 represents OR<6>and R<6>represents H or C1-C3acyl, R3 represents H orR2 and R3 together represents =0, R<4>represents OH, R<5>represents H orR <4>and R<5> together represent =0. 2. Compound according to claim 1, characterized in that R represents CH(OCH3)2, R<1> represents COCH3, R2 represents OR6 where R<6> represents H, R<3> and R<5> are equal and represent H and R<4> represents OH. 3. Compound according to claim 1, characterized in that R represents CH2N[CH2(C6H5)]2, R<1>represents COCH3, R<2>represents OR<6>where R<6>represents H, R<3>and R <5>are similar and represent H and R<4>represents OH. 4. Compound according to claim 1, characterized in that R represents CH(OCH3)2, R<1> represents COCH3, R2 represents OR6 where R6 represents COCH3, R and R are equal and represent H and R represents OH. 5. Compound according to claim 1, characterized in that R represents CH2N[CH2(C6H5)]2, R<1>represents COCH3, R<2>represents OR6 where R<6>represents COCH3, R3 and R<5> are equal and represents H and R<4>represents OH. 6. Compound according to claim 1, characterized in that R represents CH(OCH3)2, R<1>represents COCH3, R2 and R<3>together represent =0, R<4>represents OH and R<5>represents H. 7. Compound according to claim 1, characterized in that R represents CH2N[CH2(C6H5)]2, R<1> represents COCH3, R<2> and R<3> together represent =0, R<4> represents OH and R<5 >represents H. 8. Compound according to claim 1, characterized in that R represents CH(OCH3)2, R<1>represents COCH3, R<2>represents OR<6>where R<6>represents COCH3, R<3>represents H and R< 4>and R<5>together represent =0. 9. Compound according to claim 1, characterized in that R represents CH2N[CH2(C6H5)]2, R<1>represents COCH3, R2 represents OR6 where R<6>represents COCH3, R3 represents H and R<4>and R<5 >together represents =0. 10. Compound according to claim 1, characterized in that R represents CH(OCH3)2, R<1> and R<5> are equal and represent H, R<2> and R<3> together represent =0 and R<4> represent OH. 11. Compound according to claim 1, characterized in that R represents CH2N[CH2(C6H5)]2, R<1> and R5 are equal and represent H, R2 and R<3> together represent =0 and R<4> represents OH. 12. Compound according to claim 1, characterized in that R represents CH(OCH3)2, R<1>and R3 are equal and represent H, R<2>represents OR<6>where R<6>represents COCH3 and R<4>and R<5>together represents =0. 13. Compound according to claim 1, characterized in that R represents CH2N[CH2(C6H5)]2, R<1>and R3 are equal and represent H, R2 represents OR<6>where R<6>represents COCH3 and R<4>and R<5>together represents =0. 14. Compound according to claim 1, characterized in that R represents CH(OCH3)2, R<1> and R<3> are equal and represent H, R2 represents OR<6>, where R<6> represents H and R<4> and R<5>together represent =0. 15. Compound according to claim 1, characterized in that R represents CH2N[CH2(C6H5)]2, R<1>and R3 are equal and represent H, R<2>represents OR<6>where R<6>represents H and R <4>and R<5> together represent =0. 16. Compound according to claim 1, characterized in that R represents CHO, R<1> and R<3> are equal and represent H, R<2> represents OR<6>, where R6 represents H and R<4> and R<5> together represent =0. 17. Compound according to claim 1, characterized in that R represents CHO, R<1> represents COCH3, R<2> represents OR<6>, where R6 represents H, R3 and R<5> are equal and represent H and R<4> represents OH. 18. Compound according to claim 1, characterized in that R represents CHO, R<1> represents COCH3, R<2> represents OR6 where R<6> represents COCH3, R3 and R<5> are equal and represent H and R4 represents OH. 19. Compound according to claim 1, characterized in that R represents CHO, R<1> represents COCH3, R<2> and R<3> together represent =0, R<4> represents OH and R<5> represents H. 20. Compound according to claim 1, characterized in that R represents CHO, R<1> and R are equal and represent H, R and R together represent =0 and R represents OH.