CN114853782A - Pleuromutilin derivative with pyrimidine side chain and preparation method and application thereof - Google Patents

Abstract

The invention provides a pleuromutilin derivative with a pyrimidine side chain, and a preparation method and application thereof, belonging to the field of pharmaceutical chemistryThe cocci all show excellent antibacterial effect, and have low cytotoxicity and high safety.

Description

Pleuromutilin derivative with pyrimidine side chain and preparation method and application thereof

technical field

The invention belongs to the field of medicinal chemistry, and in particular relates to a pleuromutilin derivative with a pyrimidine side chain, a preparation method thereof, and use in the preparation of antibacterial drugs.

Background technique

Antibiotics have antibacterial, fungal, viral and tumor-fighting effects, and have played an important role in the history of human development since their discovery in 1928. After the 1980s, the development of antibiotics entered a slow period, and the number of newly approved antibiotics continued to decrease. In the past 40 years, only 6 antibiotics with new structures have been approved by the FDA. In addition, with the increasing antibiotic resistance, most of the antibiotics currently in use are less effective or ineffective. Therefore, there is an urgent need to develop novel antibiotics for antibacterial therapy.

Pleuromutilin was first isolated from Pleurotusmutilus (Clitopilus scyphoides) and Pleurotus passeckerianus in 1951 by KavanaghHervey and Robbins. The drug is active against Gram-positive bacteria, especially Staphylococcus aureus, with a minimum inhibitory concentration (MIC) of 0.25 μg/mL. However, due to the weak antibacterial activity and poor water solubility of pleuromutilin, its application as a drug is limited. In recent years, through molecular structure modification, pleuromutilin derivatives have achieved breakthrough research progress, and the drugs that have been used in clinical include Tiamulin, Retamorelin, Vonimulin and Azamorelin.

In the 1980s, the Sandoz Research Institute systematically studied the relationship between the structural modification of the lead compound of pleuromutilin and the antibacterial activity using the bioelectronic isosteric principle. The structure-activity relationship study showed that the tricyclic core structure of pleuromutilin, the hydroxyl group at C-11, the ester group at C-14, and the carbonyl at C-3 were all necessary functional groups for antibacterial activity. Further research found that extending the C14 side chain of pleuromutilins helps to reduce L3 protein mutation, while protecting U2584 and U2585 in the peptidyl transferase center (PTC), thereby preventing tRNA from binding to the P site. , thereby reducing the resistance to pleuromutilin compounds. In addition, pleuromutilin C14 contains a free hydroxyl group, which can penetrate into the hydrophobic part of the ribosomal subunit, resulting in its slow metabolism in vivo. By modifying the C14 position, its antibacterial activity against pathogenic bacteria (especially bacteria and mycoplasma) can be enhanced. . Therefore, modification of pleuromutilin C14 has become a research hotspot in recent years.

SUMMARY OF THE INVENTION

The object of the present invention is to provide a pleuromutilin derivative with a pyrimidine side chain and a preparation method and application thereof. The pleuromutilin derivative of the present invention has good antibacterial activity and is especially suitable for use in animals as a new type of antibacterial drug Or systemic infection of the human body.

A pleuromutilin derivative with a pyrimidine side chain is a compound shown in formula I or a stereoisomer, pharmaceutically acceptable salt or crystal form thereof:

in,

R ₁ and R ₂ are independently selected from H, (C ₁ -C ₆ )alkyl, 1-3 halogen-substituted (C ₁ -C ₆ )alkyl groups; or when R ₁ and R ₂ have no substituents at positions , R ₁ and R ₂ are connected with other atoms to form a cyclic structure A, A is selected from (C ₆ -C ₁₀ ) aryl, 5-10-membered heteroaryl, 5-10-membered heterocyclyl, the heteroaryl The base and the heterocyclyl group contain 1-3 heteroatoms selected from N, O, S, and are substituted by 0-3 identical or different R _A ;

R _A is selected from H, halogen, cyano, nitro, hydroxy, carboxyl, amino, (C ₁ -C ₆ )alkyl, 1-3 halogen or amino or hydroxy or cyano substituted (C ₁ -C ₆ ) ) alkyl, (C ₁ -C ₆ ) alkylamino, di(C ₁ -C ₆ ) alkylamino, (C ₁ -C ₆ ) alkoxy, (C ₁ -C ₆ ) alkylcarbonyl, (C 1 -C 6 ) alkylcarbonyl ₂ - _C6 )alkenyl, halo(C2 _{- C6} )alkenyl;

_{R is} selected from H, aliphatic amino,

B is selected from (C ₆ -C ₁₀ ) aryl, 5-10 membered heteroaryl, and is substituted with 0-3 identical or different R _B ;

R _B is selected from H, halogen, cyano, nitro, hydroxy, carboxyl, amino, (C ₁ -C ₆ )alkyl, 1-3 halogen or amino or hydroxy or cyano substituted (C ₁ -C ₆ ) ) alkyl, (C ₁ -C ₆ ) alkylamino, di(C ₁ -C ₆ ) alkylamino, (C ₁ -C ₆ ) alkoxy, 1-3 halogen or amino or hydroxy or cyano substitution (C ₁ -C ₆ )alkoxy, (C ₁ -C ₆ ) alkylthio, (C ₁ -C ₆ ) alkylcarbonyl, (C ₁ -C ₆ ) alkylsulfonyl, (C ₂ - C ₆ ) alkenyl, halo(C ₂ -C ₆ ) alkenyl;

Preferably, in the general formula I,

R ₁ is selected from H, (C ₁ -C ₆ )alkyl, 1-3 fluoro-substituted methyl groups;

R ₂ is selected from H, (C ₁ -C ₆ )alkyl;

Further preferably,

R ₁ is selected from H, methyl, trifluoromethyl;

R ₂ is selected from H, methyl;

Preferably, in the general formula I,

When R ₁ and R ₂ have no substituents and are connected with other atoms to form a cyclic structure A, A is selected from (C ₆ -C ₁₀ ) aryl, 4-6 membered heteroaryl, 4-6 membered heterocycle base, the heteroaryl and heterocyclyl contain 1-3 heteroatoms selected from N, O, S, and are substituted by 0-3 same or different R _A ;

R _A is selected from H, halogen, amino, (C ₁ -C ₆ )alkyl, 1-3 halogen or amino or hydroxy or cyano substituted (C ₁ -C ₆ )alkyl, (C ₁ -C ₆ )alkyl ) alkoxy, (C ₁ -C ₆ ) alkylcarbonyl, (C ₂ -C ₆ ) alkenyl;

Further preferably,

When R ₁ and R ₂ have no substituents and are connected with other atoms to form a cyclic structure A, A is selected from (C ₆ -C ₁₀ ) aryl, 4-6 membered heteroaryl, and the heteroaryl contains 1-3 heteroatoms selected from N, O, S, and substituted by 0-3 same or different R _A ;

R _A is selected from H, methyl;

More preferably,

When R ₁ and R ₂ have no substituents and are connected with other atoms to form a cyclic structure A, A is phenyl or thienyl and is substituted by R _A ;

Preferably, in the general formula I,

R ₃ is selected from H,

R _3a and R _3b are the same or different and are independently selected from H, (C ₁ -C ₆ )alkyl, (C ₂ -C ₆ )alkenyl, (C ₂ -C ₆ )alkynyl, halogen or amino or Hydroxy- or cyano-substituted (C ₁ -C ₆ )alkyl; or R _3a and R _3b together with the nitrogen atom to which they are attached form a 4-10 membered heterocyclyl group, except for R _3a and R _3b In addition to the connected nitrogen atom, it optionally contains 1-4 heteroatoms selected from N, O, S, or a sulfone group or a carbonyl group, and the heterocyclic group is optionally substituted by 0-3 identical or different R _3c ;

R _3c is selected from H, halogen, cyano, nitro, hydroxy, carboxyl, amino, (C ₁ -C ₆ )alkyl, 1-3 halogen or amino or hydroxy or cyano substituted (C ₁ -C ₆ ) ) alkyl, (C ₁ -C ₆ ) alkylamino, di(C ₁ -C ₆ ) alkylamino, (C ₁ -C ₆ ) alkoxy, (C ₁ -C ₆ ) alkylcarbonyl, (C 1 -C 6 ) alkylcarbonyl ₁ - _C6 ) alkylsulfonyl, ( _C1 - _C6 ) alkylsulfonamido, ( _C1 - _C6 ) alkylamido, (C2 _{- C6} ) alkenyl, halogenated (C2 _{) -C6} )alkenyl, alkylamino _- substituted (C2 _{- C6} )alkenyl, (C3 _- C7)cycloalkyl;

B is selected from (C ₆ -C ₁₀ ) aryl, 5-membered heteroaryl, and is substituted with 0-2 identical or different R _B ;

R _B is selected from H, halogen, nitro, hydroxy, (C ₁ -C ₆ )alkyl, 1-3 halogen substituted (C ₁ -C ₆ )alkyl, (C ₁ -C ₆ )alkoxy , (C ₁ -C ₆ ) alkylthio, (C ₁ -C ₆ ) alkylcarbonyl;

Further preferably,

R ₃ is selected from H,

R _3a and R _3b are the same or different and are independently selected from H, (C ₁ -C ₆ )alkyl, halogen or amino or hydroxy or cyano substituted (C ₁ -C ₆ )alkyl; or R _3a and R _3b together with the attached nitrogen atom forms a 5-8 membered heterocyclic group, the heterocyclic group optionally contains 0-2 selected from N, O, S in addition to the nitrogen atoms attached to R _3a and R _3b The heteroatom or sulfone group or carbonyl group, the heterocyclic group is optionally substituted by 0-2 identical or different R _3c ;

R _3c is selected from H, (C ₁ -C ₆ )alkyl, 1-3 halogen or amino or hydroxy or cyano substituted (C ₁ -C ₆ )alkyl, (C ₁ -C ₆ )alkylcarbonyl ;

More preferably,

R ₃ is selected from

The structure composed of R _3a and R _3b with the attached nitrogen atom is as follows:

B is selected from phenyl, thienyl, and is substituted by 0-2 identical or different R _B ;

R _B is selected from H, nitro, methyl, trifluoromethyl, methoxy, halogen;

Most preferably, the pleuromutilin derivative with a pyrimidine side chain according to the present invention is any one of the following compounds I-1 to I-38:

The pyrimidine side chain-containing compound represented by the general formula I of the present invention forms a pharmaceutically acceptable salt with an acid. The pharmaceutically acceptable salts include addition salts formed by inorganic and organic acids with the compounds, and the inorganic and organic acids include: hydrochloric acid, hydrogen olfactory acid, sulfuric acid, phosphoric acid, methanesulfonic acid, ethanesulfonic acid Acid, p-toluenesulfonic acid, benzenesulfonic acid, tea disulfonic acid, acetic acid, propionic acid, lactic acid, trifluoroacetic acid, maleic acid, citric acid, fumaric acid, oxalic acid, tartaric acid, benzoic acid.

In the present invention, "halogen" refers to fluorine, chlorine, bromine or iodo; "alkyl" refers to straight-chain or branched alkyl; "alkenyl" refers to straight-chain or branched alkenyl; "alkynyl" refers to "refers to straight-chain or branched alkynyl; "aryl" refers to an organic group obtained by removing a hydrogen atom in an aromatic hydrocarbon; "heteroaryl" refers to one or more selected from N, O and A heteroatom of S, wherein the ring system of each heteroaryl group may be monocyclic or polycyclic, and the ring system is aromatic; "heterocyclic group" refers to a group containing one or more selected from N, O, Monocyclic or polycyclic ring system of S heteroatom.

A pharmaceutical composition comprising the pleuromutilin derivative with a pyrimidine side chain and a pharmaceutically acceptable carrier, excipient, diluent, adjuvant, vehicle or a combination thereof.

The present invention also provides the use of the pleuromutilin derivative with a pyrimidine side chain or a pharmaceutical composition comprising the derivative in preparing an antibacterial drug. It has excellent antibacterial effect on Staphylococcus aureus, Methicillin-resistant Staphylococcus aureus and Streptococcus agalactiae.

The pleuromutilin derivative with pyrimidine side chain in the present invention has novel structure and excellent antibacterial activity. It can be clearly seen from the in vitro antibacterial experiments that it has excellent antibacterial effects against Staphylococcus aureus, methicillin-resistant Staphylococcus aureus, and Streptococcus agalactiae, with low cytotoxicity and high safety, and is expected to be used in the treatment of Bacterial infections caused by Staphylococcus aureus and Streptococcus agalactiae provide new options for anti-infective drugs.

The present invention also provides a method for preparing the pyrimidine side chain-containing pleuromutilin derivative represented by general formula I.

The synthetic route of the present invention comprises the steps:

Synthetic route 1:

时，中间体C-1的制备方法如下，该中间体用于制备化合物I-1～I-26；When R in intermediate D is

, the preparation method of intermediate C-1 is as follows, and this intermediate is used to prepare compounds I-1～I-26;

Synthetic route 2:

时，中间体C-2的制备方法如下，该中间体用于制备化合物I-37～I-38；When R in intermediate D is

, the preparation method of intermediate C-2 is as follows, and this intermediate is used to prepare compounds I-37～I-38;

Synthetic route 3:

时，中间体C采用化合物

(2-巯基喹唑林-4-醇)，该中间体用于制备化合物I-27～I-28；当中间体D中的R为

时，中间体C采用化合物

该中间体用于制备化合物I-29～I-30；当中间体D中的R为

时，中间体C采用化合物

该中间体用于制备化合物I-31～I-32；当中间体D中的R为

时，中间体C采用化合物

该中间体用于制备化合物I-33～I-34；当中间体D中的R为

时，中间体C采用化合物

该中间体用于制备化合物I-35～I-36。When R in intermediate D is

, intermediate C adopts compound

(2-mercaptoquinazolin-4-ol), this intermediate is used to prepare compounds I-27～I-28; when R in intermediate D is

, intermediate C adopts compound

This intermediate is used to prepare compounds I-29～I-30; when R in intermediate D is

, intermediate C adopts compound

This intermediate is used to prepare compounds I-31～I-32; when R in intermediate D is

, intermediate C adopts compound

This intermediate is used to prepare compounds I-33～I-34; when R in intermediate D is

, intermediate C adopts compound

This intermediate is used to prepare compounds I-35 to I-36.

Beneficial effects of the present invention:

The compound of the invention has a novel chemical structure, and has excellent antibacterial effect against Staphylococcus aureus, methicillin-resistant Staphylococcus aureus and Streptococcus agalactiae in in vitro studies, and has low cytotoxicity and high safety. .

Description of drawings

Figure 1 Time-killing curves of different concentrations of compounds I-1, I-16 and Tiamulin on S. aureus.

Detailed ways

In the following examples, methods for preparing compounds of formula I are provided. It will be appreciated that the following methods and other methods known to those of ordinary skill in the art can be applied to the preparation of all compounds described herein. The examples are intended to illustrate rather than limit the scope of the invention.

Example 1: 14-O-[(4-(4-ethylpiperazin-1-yl)thieno[2,3-d]pyrimidin-2-yl)mercaptoacetyl]mutilin (I-1 ), prepared by the method of synthetic route 1

1.1: Preparation of 14-O-(p-toluenesulfonyloxyacetyl)-mutilin (Intermediate B)

At -20°C, pleuromutilin (10 g, 26.4 mmol) and water (15 mL) were added to methyl tert-butyl ether (30 mL), followed by p-toluenesulfonyl chloride (5.54 g, 29.05 mmol) and sodium hydroxide (1 g, 25 mmol), and after stirring for 15 min, the reaction was heated to 55 °C for 3 h. After the reaction was completed, the reaction solution was cooled to room temperature, 20 mL of ice-water mixture was added, stirred for 15 min, suction filtered, the filter cake was washed with a small amount of clear water, and dried to obtain a crude product. The crude product was washed with petroleum ether, filtered and dried to obtain 10.7 g of a white solid, yield: 74.5%.

1.2: 14-O-(p-toluenesulfonyloxyacetyl)-mutilin (intermediate C-1), prepared by the method of synthetic route 2

1.2.1: Preparation of ethyl 2-(3-benzoylthiourea)thiophene-3-carboxylate (Intermediate H)

Benzoyl chloride (15.9 g, 113.15 mmol) and ammonium thiocyanate (12.9 g, 169.47 mmol) were added to dry acetonitrile (30 mL) solution at room temperature, and the reaction solution was warmed to room temperature and stirred under nitrogen system The reaction was carried out for 30 minutes. At this temperature, 2-aminothiophene-3-carboxylic acid ethyl ester (10 g, 56.42 mmol) was added, and the reaction was continued to stir for 6 h. After the reaction was completed, the reaction solution was cooled to 0° C. in an ice bath, filtered, and the filter cake was washed with cold acetonitrile and water successively, and the filter cake was dried to obtain 18.9 g of a yellow solid, yield: 96.9%.

1.2.2: 14-O-(p-toluenesulfonyloxyacetyl)-mutilin (Intermediate C-1)

Intermediate H (23.5 g, 70.35 mmol) and potassium hydroxide (22.45 g, 401.1 mmol) were added to ethanol (50 mL), and the reaction solution was heated to 82 °C and stirred for 19 h. After the reaction was completed, the reaction solution was cooled to room temperature, the solvent was evaporated, the pH was acidified to 7 with 1N hydrochloric acid, the solid was precipitated, filtered with suction, the filter cake was stirred and washed with a small amount of ethanol, and the filter cake was dried to obtain 12.1 g of white solid with a yield of 94.0%.

1.3: Preparation of 14-O-[(4-hydroxythieno[2,3-d]pyrimidin-2-yl)mercaptoacetyl]mutilin (Intermediate D)

Intermediate B (12.275 g, 23.06 mmol), Intermediate C-1 (4.25 g, 23.1 mmol) and sodium hydroxide (1.22 g) were added to 1:1 methanol/methyl tert-butyl ether (30 mL), The reaction was stirred at 65 °C for 6 h. After the reaction was completed, the solvent was evaporated, added to 100 mL of water, extracted with dichloromethane (30 mL × 3), the organic layers were combined, dried over anhydrous sodium sulfate, filtered with suction, and evaporated to dryness under reduced pressure to obtain 8.3 g of white solid with a yield of 65.8 g %.

1.4: 14-O-[(4-(2,4,6-trimethylbenzenesulfonate-1-yl)thieno[2,3-d]pyrimidin-2-yl)mercaptoacetyl]mutilin

Preparation of (Intermediate E)

Intermediate D (8.26 g, 15.18 mmol), triethylamine (2.3 g, 22.72 mmol) and 4-dimethylaminopyridine (0.826 g, 10 mmol) were added to N,N-dimethylmethane under ice bath conditions amide (30 mL), 2,4,6-trimethylbenzenesulfonyl chloride (4.98 g, 22.72 mmol) was slowly added thereto, and the reaction was stirred at 0° C. for 1 h. After the reaction was completed, 100 mL of water was added, extracted with dichloromethane (30 mL×3), the organic layers were combined, dried over anhydrous sodium sulfate, filtered with suction and evaporated to dryness under reduced pressure to obtain 10.7 g of a white solid with a yield of 97.0%.

1.5: 14-O-[(4-(4-ethylpiperazin-1-yl)thieno[2,3-d]pyrimidin-2-yl)mercaptoacetyl]mutilin (I-1) Preparation Intermediate E (0.3 g, 0.413 mmol), triethylamine (0.42 g, 4.15 mmol), DMAP (0.01 g, 0.082 mmol) and 1-ethylpiperazine (0.24 g, 2.065 mmol) were prepared under ice bath conditions mmol) was added to DMF (8 mL), and the reaction was stirred at 0 °C for 1-2 h. After the reaction was completed, the solvent was evaporated, and the crude product obtained was purified by silica gel column using petroleum ether/ethyl acetate (3:1 to 1:1, v/v) as the eluent to obtain 0.1 g of a light white solid with a yield of 37.6%.

mp: 115.6-116.1°C; ¹ H-NMR (400 MHz, CDCl ₃ ) δ 7.24 (d, J=6.1 Hz, 1H), 7.11 (d, J=6.1 Hz, 1H), 6.47 (dd, J=17.2 ,11.0Hz,1H),5.75(dd,J＝15.4,8.4Hz,1H),5.28(d,J＝11.0Hz,1H),5.14(d,J＝17.4Hz,1H),3.99(s,3H ), 3.95–3.83 (m, 2H), 3.29 (d, J=36.0Hz, 2H), 2.69–2.49 (m, 6H), 2.35–2.17 (m, 4H), 2.08 (s, 1H), 2.01 ( dd, J=15.9, 8.5Hz, 1H), 1.64 (td, J=44.7, 43.1, 13.5Hz, 5H), 1.45 (s, 2H), 1.41–1.24 (m, 4H), 1.18 (t, J= 7.3Hz, 3H), 1.12(s, 3H), 0.85(d, J=6.7Hz, 3H), 0.76(d, J=6.9Hz, 3H). ¹³ C-NMR (101MHz, CDCl ₃ )δ217.16,170.15 ,168.35,157.74,138.97,120.33,120.08,117.16,113.47,69.60,58.17,52.38,46.41,45.45,44.59,43.99,41.90,36.83,35.96,34.50,34.31,30.45,26.91,26.32,24.82,16.98,14.94 ,11.54,11.46.HRMS(ES)calcd[M+H] ⁺ for C ₃₄ H ₄₈ N ₄ O ₄ S ₂ 641.3195,found 641.3196.

Example 2: 14-O-[(4-((2-(dimethylamino)ethyl)(methyl)amino)thieno[2,3-d]pyrimidin-2-yl)mercaptoacetyl] Preparation of mutilin (I-2)

According to the synthesis method of Example 1, the compound I-2 of Example 2 is obtained by reacting intermediate E with N ¹ , N ¹ , N ² -trimethylethane-1,2-diamine.

HRMS(ES)calcd[M+H] ⁺ for C ₃₃ H ₄₈ N ₄ O ₄ S ₂ 629.3195, found 629.3196.

Example 3: 14-O-[(4-(4-Acetylpiperazin-1-yl)thieno[2,3-d]pyrimidin-2-yl)mercaptoacetyl]mutilin (I-3 ) preparation

According to the synthetic method of Example 1, the compound I-3 of Example 3 was obtained by reacting intermediate E with 1-(piperazin-1-yl)ethane-1-one.

HRMS(ES)calcd[M+H] ⁺ for C ₃₃ H ₄₈ N ₄ O ₄ S ₂ 655.2982, found 655.2980.

Example 4: 14-O-[(4-(4-methylpiperidin-1-yl)thieno[2,3-d]pyrimidin-2-yl)mercaptoacetyl]mutilin (I-4 ) preparation

According to the synthetic method of Example 1, the compound I-4 of Example 4 is obtained by reacting intermediate E with 4-methylpiperidine.

mp: 89.6-90.2°C; ¹ H-NMR (400 MHz, CDCl ₃ ) δ 7.23 (d, J=5.9 Hz, 1H), 7.07 (d, J=5.9 Hz, 1H), 6.48 (dd, J=17.3 ,11.0Hz,1H),5.73(d,J＝8.2Hz,1H),5.28(d,J＝10.8Hz,1H),5.14(d,J＝17.3Hz,1H),4.56(s,2H), 3.90(d,J＝5.2Hz,2H),3.43(d,J＝12.6Hz,1H),3.33(d,J＝5.4Hz,1H),3.11-3.04(m,2H),2.90-2.80(m ,1H),2.33–2.29(m,1H),2.24–2.17(m,2H),2.10–1.97(m,3H),1.80(s,1H),1.76(s,1H),1.73(s,1H) ),1.65–1.59(m,3H),1.54(d,J=13.7Hz,1H),1.47(d,J=10.5Hz,1H),1.36(d,J=7.9Hz,2H),1.30(d , J＝5.4Hz, 2H), 1.26(s, 2H), 1.12(s, 3H), 0.99(d, J＝6.0Hz, 3H), 0.85(d, J＝6.6Hz, 3H), 0.76(d , J=6.5Hz, 3H). ¹³ C-NMR (101MHz, CDCl ₃ )δ168.55,155.37,139.16,120.86,118.58,116.98,113.34,69.41,58.18,53.75,48.86,45.45,44.48,43.94,41.9 ,36.65,35.97,34.49,34.31,30.45,26.90,26.27,24.82,23.97,16.90,14.93,11.45.HRMS(ES)calcd[M+H] ⁺ for C ₃₄ H ₄₇ N ₃ O ₄ S ₂ 626.3086,found 626.3087.

Example 5: Preparation of 14-O-[(4-morpholinthieno[2,3-d]pyrimidin-2-yl)mercaptoacetyl]mutilin (I-5)

According to the synthetic method of Example 1, the compound I-5 of Example 5 is obtained by reacting intermediate E with morpholine.

HRMS(ES)calcd[M+H] ⁺ for C ₃₆ H ₅₃ N ₄ O ₆ S ₂ 701.3407, found 701.3405.

Example 6: 14-O-[(4-(4-(2-hydroxyethyl)piperazin-1-yl)thieno[2,3-d]pyrimidin-2-yl)mercaptoacetyl]mer Preparation of Lin (I-6)

According to the synthetic method of Example 1, the compound I-6 of Example 6 was obtained by reacting intermediate E with 2-(piperazin-1-yl)ethane-1-ol.

HRMS(ES)calcd[M+H] ⁺ for C ₃₆ H ₅₃ N ₄ O ₆ S ₂ 657.3139, found 657.3138.

Example 7: Preparation of 14-O-[(4-(piperazin-1-yl)thieno[2,3-d]pyrimidin-2-yl)mercaptoacetyl]mutilin (I-7)

According to the synthetic method of Example 1, the compound I-7 of Example 7 is obtained by reacting intermediate E with piperazine.

HRMS(ES)calcd[M+H] ⁺ for C ₃₂ H ₄₄ N ₄ O ₄ S ₂ 613.2882, found 613.2889.

Example 8: Preparation of 14-O-[(4-(dimethylamino)thieno[2,3-d]pyrimidin-2-yl)mercaptoacetyl]mutilin (I-8)

According to the synthetic method of Example 1, the compound I-8 of Example 8 is obtained by reacting intermediate E with dimethylamine.

HRMS(ES)calcd[M+H] ⁺ for C ₃₀ H ₄₂ N ₃ O ₄ S ₂ 572.2617, found 572.2618.

Example 9: Preparation of 14-O-[(4-(diethylamino)thieno[2,3-d]pyrimidin-2-yl)mercaptoacetyl]mutilin (I-9)

According to the synthetic method of Example 1, the compound I-9 of Example 9 is obtained by reacting intermediate E with diethylamine.

HRMS(ES)calcd[M+H] ⁺ for C ₃₀ H ₄₂ N ₃ O ₄ S ₂ 600.2924, found 600.2922.

Example 10: Preparation of 14-O-[(4-thiomorpholinethieno[2,3-d]pyrimidin-2-yl)mercaptoacetyl]mutilin (I-10)

According to the synthetic method of Example 1, the compound I-10 of Example 10 is obtained by reacting intermediate E with thiomorpholine.

HRMS(ES)calcd[M+H] ⁺ for C ₃₂ H ₄₃ N ₃ O ₄ S ₃ 630.2494, found 630.2498.

Example 11: Preparation of 14-O-[(4-(piperidin-1-yl)thieno[2,3-d]pyrimidin-2-yl)mercaptoacetyl]mutilin (I-11)

According to the synthetic method of Example 1, the compound I-11 of Example 11 was obtained by reacting intermediate E with piperidine.

HRMS(ES)calcd[M+H] ⁺ for C ₃₂ H ₄₃ N ₃ O ₄ S ₃ 612.2924, found 612.2921.

Example 12: Preparation of 14-O-[(4-(pyrrolidin-1-yl)thieno[2,3-d]pyrimidin-2-yl)mercaptoacetyl]mutilin (I-12)

According to the synthetic method of Example 1, the compound I-12 of Example 12 is obtained by reacting intermediate E with pyrrolidine.

HRMS(ES)calcd[M+H] ⁺ for C ₃₂ H ₄₄ N ₃ O ₄ S ₂ 598.2773, found 598.2775.

Example 13: 14-O-[(4-(4-(Dimethylamino)piperidin-1-yl)thieno[2,3-d]pyrimidin-2-yl)mercaptoacetyl]mutilin Preparation of (I-13)

According to the synthetic method of Example 1, the compound I-13 of Example 13 was obtained by reacting intermediate E with N,N-dimethylpiperidin-4-amine.

mp: 133.0-134.1°C; ¹ H-NMR (400MHz, CDCl ₃ ) δ 7.22 (d, J=6.0 Hz, 1H), 7.15 (d, J=5.9 Hz, 1H), 6.95 (s, 1H), 6.82(s, 1H), 6.43(dd, J=17.4, 10.9Hz, 1H), 5.70(d, J=8.3Hz, 1H), 5.23(d, J=10.9Hz, 1H), 3.87(d, J =11.4Hz,1H),3.73(d,J=12.8Hz,1H),3.35(d,J=5.5Hz,2H),3.14(s,1H),3.12(s,1H),2.81(s,3H ), 2.77(s, 3H), 2.63(s, 4H), 2.56(s, 3H), 2.30(s, 3H), 2.22(s, 3H), 2.10(s, 1H), 1.77(d, J= 11.6Hz, 3H), 1.63(s, 1H), 1.38(s, 1H), 1.36(s, 2H), 1.35(s, 1H), 1.12(s, 3H), 0.85(d, J=6.7Hz, 3H), 0.75(d, J=6.8Hz, 3H). ¹³ C-NMR (101MHz, CDCl ₃ )δ168.20, 140.33, 139.14, 136.83, 132.11, 130.78, 120.73, 116.95, 69.76, 63.11, 62.70, 58.13, 46.03 ,45.43,44.00,43.08,41.88,39.69,36.78,35.98,34.50,30.40,26.54,25.65,24.78,22.98,22.78,20.97,20.74,16.93,14.91,11.46.HRMS] ⁺ HRMS(ES)calcd[M+HRMS(ES)] for C ₃₅ H ₅₁ N ₄ O ₄ S ₂ 655.3352, found 655.3355.

Example 14: 14-O-[(4-(4-Methyl-1,4-diazepan-1-yl)thiophene[2,3-d]pyrimidin-2-yl)mercaptoacetyl ] Preparation of mutilin (I-14)

According to the synthetic method of Example 1, the compound I-14 of Example 14 was obtained by reacting intermediate E with 1-methyl-1,4-diazepane.

HRMS(ES)calcd[M+H] ⁺ for C ₄₀ H ₆₃ N ₆ O ₄ S ₂ 755.4352, found 755.4355.

Example 15: 14-O-[(4-(4-methylpiperazin-1-yl)thieno[2,3-d]pyrimidin-2-yl)mercaptoacetyl]mutilin (I-15 ) preparation

According to the synthetic method of Example 1, the compound I-15 of Example 15 was obtained by reacting intermediate E with 4-methylpiperazine.

HRMS(ES)calcd[M+H] ⁺ for C ₃₃ H ₄₇ N ₄ O ₄ S ₂ 627.3039, found 627.3038.

Example 16: Preparation of 14-O-[(4-((2-hydroxyethyl)amino)thieno[2,3-d]pyrimidin-2-yl]mutilin (I-16)

According to the synthetic method of Example 1, the compound I-16 of Example 16 was obtained by reacting intermediate E with 2-aminoethyl-1-ol.

HRMS(ES)calcd[M+H] ⁺ for C ₃₀ H ₄₁ N ₃ O ₅ S ₂ 588.2566, found 588.2568.

According to the method of route 1, intermediate E prepared in Example 1 is reacted with hydrazine hydrate to obtain intermediate F, and intermediate F is reacted with formaldehyde of different substitutions to obtain compounds I-17～I-26 of embodiment 17-26 .

Example 17: Preparation of 14-O-[(4-(2-benzylidenehydrazine)thieno[2,3-d]pyrimidin-2-yl)mercaptoacetyl]mutilin (I-17)

1.1: Preparation of 14-O-[(4-hydrazinothieno[2,3-d]pyrimidin-2-yl)mercaptoacetyl]mutilin (Intermediate F)

Under ice bath conditions, intermediate E (5 g, 7 mmol) prepared in Example 1 and hydrazine hydrate (4.15 g, 35 mmol) were added to dichloromethane (15 mL), and the reaction was stirred at 0 °C for 2 h. After the reaction was completed, the solvent was evaporated to obtain 3.7 g of a white solid with a yield of 96.2%.

1.2: Preparation of 14-O-[(4-(2-benzylidenehydrazine)thieno[2,3-d]pyrimidin-2-yl)mercaptoacetyl]mutilin (I-17)

Combine 14-O-[(4-hydrazinothieno[2,3-d]pyrimidin-2-yl)mercaptoacetyl]magtiline (0.3 g, 0.54 mmol) and benzaldehyde (0.06 g, 0.594 mmol) It was added to isopropanol (5 mL), and the reaction was stirred at room temperature for 4 h. The solvent was evaporated, and the crude product obtained was purified by silica gel column using petroleum ether/ethyl acetate (3:1 to 1:1, v/v) as eluent to obtain 0.13 g of a light white solid with a yield of 38.4%.

HRMS(ES)calcd[M+H] ⁺ for C ₃₂ H ₄₃ N ₃ O ₄ S ₃ 647.2720, found 647.2722.

Example 18: 14-O-[(4-(2-(4-(trifluoromethyl)benzylidene)hydrazino)thieno[2,3-d]pyrimidin-2-yl)mercaptoacetyl ] Preparation of mutilin (I-18)

According to the synthetic method of Example 17, intermediate F was reacted with 4-(trifluoromethyl)benzaldehyde to obtain compound I-18 of Example 18.

mp: 111.4-112.7°C; ¹ H-NMR (400 MHz, CDCl ₃ ) δ 9.30 (s, 1H), 7.99 (d, J=5.9 Hz, 1H), 7.93 (s, 1H), 7.81 (d, J =8.0Hz,2H),7.70(d,J=8.1Hz,2H),6.53-6.47(m,1H),5.75(d,J=8.3Hz,1H),5.39-5.33(m,1H),5.28 (d, J=10.9Hz, 1H), 5.14(d, J=17.5Hz, 1H), 3.95(s, 1H), 3.85(s, 1H), 3.43(s, 1H), 2.21(d, J= 7.2Hz, 3H), 2.11(s, 1H), 2.08(s, 1H), 1.77(s, 1H), 1.73(s, 1H), 1.65(s, 1H), 1.62(s, 1H), 1.40( s,2H),1.36(s,1H),1.33(s,1H),1.29(s,1H),1.25(s,1H),1.17(s,1H),1.16(s,1H),1.10(s , 3H), 0.85(d, J=6.9Hz, 3H), 0.80(d, J=6.8Hz, 3H). ¹³ C-NMR (101MHz, CDCl ₃ )δ217.10, 168.22, 154.73, 142.12, 139.44, 139.13, 137.21,127.10,125.94,125.91,122.29,121.70,117.12,112.82,69.78,58.15,45.45,44.46,43.96,41.90,36.81,35.98,34.47,30.43,26.92,26.85,26.25,24.81,16.99,14.93,11.45. HRMS(ES)calcd[M+H] ⁺ for C ₃₆ H ₄₅ F ₃ N ₄ O ₄ S ₂ 715.2600, found 715.2601.

Example 19: 14-O-[(4-(2-(2-Methoxybenzylidene)hydrazino)thieno[2,3-d]pyrimidin-2-yl)mercaptoacetyl]mer Preparation of Lin(I-19)

According to the synthetic method of Example 17, intermediate F was reacted with 2-(methoxy)benzaldehyde to obtain compound I-19 of Example 19.

HRMS(ES)calcd[M+H] ⁺ for C ₃₆ H ₄₅ N ₄ O ₅ S ₂ 677.2831, found 677.2833.

Example 20: 14-O-[(4-(2-(4-Bromo-3-nitrophenyl)hydrazino)thieno[2,3-d]pyrimidin-2-yl)mercaptoacetyl]mu Preparation of Tilin (I-20)

According to the synthetic method of Example 17, intermediate F was reacted with 4-bromo-3-nitrobenzaldehyde to obtain compound I-20 of Example 20.

HRMS(ES)calcd[M+H] ⁺ for C ₃₆ H ₄₅ N ₄ O ₅ S ₂ 677.2831, found 677.2833.

Example 21: 14-O-[(4-(2-(4-fluorobenzylidene)hydrazino)thieno[2,3-d]pyrimidin-2-yl)mercaptoacetyl]mutilin (I -21) Preparation of

According to the synthetic method of Example 17, the compound I-21 of Example 21 was obtained by reacting intermediate F with 4-fluorobenzaldehyde.

¹ H-NMR (400 MHz, CDCl ₃ ) δ 7.99 (d, J=6.0 Hz, 1H), 7.86 (s, 1H), 7.73-7.65 (m, 2H), 7.22 (d, J=6.0 Hz, 1H) ), 7.15(t, J＝8.3Hz, 2H), 6.51(dd, J＝17.4, 11.1Hz, 1H), 5.75(d, J＝8.4Hz, 1H), 5.29(d, J＝11.0Hz, 1H) ), 5.14 (d, J＝17.5Hz, 1H), 3.96 (d, J＝16.5Hz, 1H), 3.82 (d, J＝16.6Hz, 1H), 3.36–3.31 (m, 1H), 2.33–2.29 (m, 1H), 2.25–2.19 (m, 2H), 2.08 (s, 1H), 2.02–1.96 (m, 1H), 1.75 (d, J=13.4Hz, 2H), 1.66 (d, J=11.6 Hz, 2H), 1.61(d, J＝10.8Hz, 1H), 1.54(d, J＝14.0Hz, 1H), 1.41–1.33(m, 3H), 1.30(s, 1H), 1.25(s, 1H ),1.18(s,1H),1.12(d,J＝4.1Hz,1H),1.10(s,3H),0.85(d,J＝7.0Hz,3H),0.80(d,J＝6.9Hz,3H ).HRMS(ES)calcd[M+H] ⁺ for C ₃₅ H ₄₂ FN ₄ O ₄ S ₂ 665.2632, found 665.2632.

Example 22: 14-O-[(4-(2-(2,4-Difluorobenzylidene)hydrazine)thieno[2,3-d]pyrimidin-2-yl)mercaptoacetyl]mutilin Preparation of (I-22)

According to the synthetic method of Example 17, the compound I-22 of Example 22 was obtained by reacting intermediate F with 2,4-difluorobenzaldehyde.

HRMS(ES)calcd[M+H] ⁺ for C ₃₅ H ₄₀ F ₂ N ₄ O ₄ S ₂ 683.2537, found 683.2535.

Example 23: 14-O-[(4-(2-(4-Methoxybenzylidene)hydrazino)thieno[2,3-d]pyrimidin-2-yl)mercaptoacetyl]mer Preparation of Lin (I-23)

According to the synthetic method of Example 17, the compound I-23 of Example 23 was obtained by reacting intermediate F with 4-methoxybenzaldehyde.

HRMS(ES)calcd[M+H] ⁺ for C ₃₆ H ₄₄ N ₄ O ₅ S ₂ 677.2831, found 677.2830.

Example 24: 14-O-[(4-(2-(2-Nitrobenzylidene)hydrazino)thieno[2,3-d]pyrimidin-2-yl)mercaptoacetyl]mutilin Preparation of (I-24)

According to the synthetic method of Example 17, the compound I-24 of Example 24 was obtained by reacting intermediate F with 2-nitrobenzaldehyde.

mp: 89.6-90.4°C; ¹ H-NMR (400 MHz, CDCl ₃ ) δ 9.22 (s, 1H), 8.50 (s, 1H), 8.15 (d, J=7.8 Hz, 1H), 8.10 (d, J =8.2Hz,1H),7.91(d,J=5.9Hz,1H),7.74(t,J=7.6Hz,1H),7.57(t,J=7.8Hz,1H),7.23(d,J=5.9 Hz, 1H), 6.55–6.48 (m, 1H), 5.76 (d, J=8.5Hz, 1H), 5.32 (d, J=10.9Hz, 1H), 5.16 (d, J=17.4Hz, 1H), 3.97(d,J=16.5Hz,1H),3.84-3.79(m,1H),3.34(d,J=12.1Hz,1H),2.34-2.29(m,1H),2.24-2.20(m,2H) ,2.08(s,1H),2.03–1.97(m,1H),1.75(d,J＝14.3Hz,2H),1.66(d,J＝9.8Hz,3H),1.41(s,1H),1.32( s, 1H), 1.28(s, 1H), 1.25(s, 1H), 1.18(s, 1H), 1.16(s, 1H), 1.13–1.11(m, 1H), 1.09(s, 3H), 0.85 (d, J=7.0Hz, 3H), 0.80 (d, J=6.9Hz, 3H). ¹³ C-NMR (101MHz, CDCl ₃ ) δ 217.08, 147.87, 139.58, 139.16, 135.90, 133.66, 130.15, 128.95, 128.22 ,125.20,122.00,121.88,117.23,69.72,58.15,45.45,44.45,43.94,41.89,36.81,35.98,34.47,34.39,30.43,26.92,26.22,24.82,17.00,14.94 +H] ⁺ for C ₃₅ H ₄₂ N ₅ O ₆ S ₂ 692.2577, found 692.2578.

Example 25: 14-O-[(4-(2-(4-Methyl-2-nitrophenyl)hydrazino)thieno[2,3-d]pyrimidin-2-yl)mercaptoacetyl] Preparation of Mutilin (I-25)

According to the synthetic method of Example 17, the compound I-25 of Example 25 was obtained by reacting intermediate F with 4-methyl-2-nitrobenzaldehyde.

HRMS(ES)calcd[M+H] ⁺ for C ₃₅ H ₄₂ N ₅ O ₆ S ₂ 706.2728, found 706.2726.

Example 26: 14-O-[(4-(2-(Thien-2-ylmethylene)hydrazino)thieno[2,3-d]pyrimidin-2-yl)mercaptoacetyl]mutilin Preparation of (I-26)

According to the synthetic method of Example 17, intermediate F was reacted with thiophene-2-carbaldehyde to obtain compound I-26 of Example 26.

mp: 132.8–133.9°C; ¹ H-NMR (400 MHz, CDCl ₃ ) δ 9.15 (s, 1H), 8.05 (s, 1H), 8.01 (d, J=5.6 Hz, 1H), 7.39 (d, J ＝4.2Hz, 1H), 7.20(d, J＝5.9Hz, 1H), 7.08(s, 1H), 6.50(dd, J＝17.4, 10.9Hz, 1H), 5.75(d, J＝8.0Hz, 1H) ), 5.29(d, J＝11.0Hz, 1H), 5.14(d, J＝17.5Hz, 1H), 3.95(d, J＝16.3Hz, 1H), 3.81(d, J＝15.9Hz, 1H), 3.38(d,J＝38.3Hz,2H),2.33-2.29(m,1H),2.21(d,J＝6.1Hz,2H),2.08(s,1H),1.99(d,J＝8.1Hz,1H ), 1.77(s, 1H), 1.73(s, 1H), 1.67(s, 1H), 1.63(d, J=10.6Hz, 2H), 1.35(d, J=12.0Hz, 3H), 1.27(d , J＝15.0Hz, 2H), 1.17(d, J＝7.9Hz, 2H), 1.09(s, 3H), 0.85(d, J＝6.6Hz, 3H), 0.79(d, J＝6.7Hz, 3H) ).HRMS(ES)calcd[M+H] ⁺ for C ₃₃ H ₄₁ N ₄ O ₄ S ₃ 653.2290, found 653.2294.

According to the synthesis method of Example 1, when the intermediate C is 2-mercaptoquinazolin-4-ol, the intermediate E is reacted with different small-molecule fatty amines to obtain the compounds I-27 to I-28 of Examples 27-28 .

Example 27: Preparation of 14-O-[(4-(4-ethylpiperazin-1-yl)quinazolin-2-yl)mercaptoacetyl]mutilin (I-27)

According to the synthetic method of Example 1, the compound I-27 of Example 27 was obtained by reacting intermediate E with 1-ethylpiperazine.

mp: 104.4-106.0°C; ¹ H-NMR (400 MHz, CDCl ₃ ) δ 7.77 (d, J=8.2 Hz, 1H), 7.64 (q, J=8.3 Hz, 2H), 7.31 (t, J=7.2 Hz,1H),6.47(dd,J=17.3,11.0Hz,1H),5.73(d,J=8.3Hz,1H),5.26(d,J=10.9Hz,1H),5.12(d,J=17.4 Hz, 1H), 3.95(d, J＝10.7Hz, 2H), 3.85(s, 4H), 3.31(s, 1H), 2.67(s, 4H), 2.55(q, J＝6.9Hz, 2H), 2.33–2.28(m, 1H), 2.20(t, J=11.9Hz, 2H), 2.06(s, 1H), 1.97(dd, J=15.8, 8.7Hz, 2H), 1.74(d, J=14.5Hz) ,1H),1.64(s,1H),1.60(d,J＝10.0Hz,2H),1.53(d,J＝12.8Hz,1H),1.45(s,2H),1.34(d,J＝13.2Hz ,2H),1.27(d,J＝12.2Hz,1H),1.17(t,J＝7.1Hz,3H),1.11(d,J＝3.8Hz,1H),1.07(s,3H),0.85(d , J=6.8Hz, 3H), 0.75 (d, J=6.7Hz, 3H). ¹³ C-NMR (101MHz, CDCl ₃ )δ217.13, 168.48, 163.65, 152.51, 138.98, 132.69, 127.50, 125.11, 124.08, 117.12 ,114.23,69.47,58.17,52.48,52.38,49.28,45.44,44.51,43.92,41.89,36.82,35.95,34.48,34.33,34.30,30.44,26.89,26.23,24.81,16.9ES2,14.94(115.9ES2,146.94 )calcd[M+H] ⁺ for C ₃₆ H ₅₀ N ₄ O ₄ S635.3631, found 635.3632.

Example 28: Preparation of 14-O-[(4-((2-hydroxyethyl)amino)quinazolin-2-yl)mercaptoacetyl]mutilin (I-28)

According to the synthetic method of Example 1, the compound I-28 of Example 28 was obtained by reacting intermediate E with 2-aminoethyl-1-ol.

¹ H-NMR (400 MHz, CDCl ₃ ) δ 7.69 (d, J=8.2 Hz, 1H), 7.63-7.56 (m, 2H), 7.31 (t, J=7.4 Hz, 1H), 6.58-6.43 (m ,2H),5.74(d,J＝8.5Hz,1H),5.26(d,J＝11.0Hz,1H),5.12(d,J＝17.4Hz,1H),3.97(d,J＝16.2Hz,1H) ),3.90(d,J＝7.8Hz,3H),3.85(s,1H),3.78(d,J＝5.3Hz,1H),3.32(d,J＝5.0Hz,1H),2.34–2.26(m , 2H), 2.25–2.19 (m, 2H), 2.18–2.11 (m, 1H), 2.10–1.93 (m, 3H), 1.75 (d, J=14.3Hz, 1H), 1.63 (q, J=11.7 ,11.3Hz,3H),1.53(d,J＝13.8Hz,1H),1.41-1.33(m,2H),1.28(d,J＝19.0Hz,1H),1.20-0.97(m,5H),0.85 (d, J=6.9Hz, 3H), 0.76 (d, J=6.9Hz, 3H).HRMS(ES) calcd[M+H] ⁺ for C ₃₂ H ₄₃ N ₃ O ₅ S 582.3001, found582.3005.

According to the synthesis method of Example 1, when the intermediate C is 2-mercapto-6-methylpyrimidin-4-ol, the intermediate E reacts with different small-molecule fatty amines to obtain the compounds I-29 of Examples 29-30 to I-30.

Example 29: Preparation of 14-O-[(4-(4-ethylpiperazin-1-yl)-6-methylpyrimidin-2-yl)mercaptoacetyl]mutilin (I-29)

According to the synthetic method of Example 1, the compound I-29 of Example 29 was obtained by reacting intermediate E with 1-ethylpiperazine.

mp: 127.8-128.7°C; ¹ H-NMR (400 MHz, CDCl ₃ ) δ 6.49 (dd, J=17.3, 11.0 Hz, 1H), 6.05 (s, 1H), 5.71 (d, J=8.4 Hz, 1H) ),5.32(d,J＝11.0Hz,1H),5.17(d,J＝17.4Hz,1H),3.83(s,2H),3.70(s,4H),3.34(dd,J＝9.9,6.6Hz ,1H),2.56(d,J＝15.7Hz,6H),2.28(d,J＝17.5Hz,5H),2.19(dd,J＝18.4,9.0Hz,2H),2.10–1.96(m,3H) ,1.75(d,J＝14.1Hz,1H),1.67(d,J＝12.1Hz,1H),1.63(s,2H),1.50(dd,J＝20.0,12.4Hz,3H),1.33(d, J=22.6Hz, 2H), 1.26(s, 1H), 1.18(t, J=7.1Hz, 3H), 1.13(s, 3H), 0.86(d, J=6.8Hz, 3H), 0.71(d, J=6.7Hz, 3H). ¹³ C-NMR (101MHz, CDCl ₃ )δ217.11,168.45,165.92,139.11,117.12,97.53,69.49,58.17,52.37,52.12,45.45,44.59,43.96,43.41,41.90,36 35.96,34.49,34.09,30.43,26.90,26.36,24.80,24.14,16.86,14.93,11.49,11.44.HRMS(ES)calcd[M+H] ⁺ for C ₃₃ H ₅₀ N ₄ O ₄ S 599.3631, found 599.3631.

Example 30: Preparation of 14-O-[(4-((2-hydroxyethyl)amino)-6-methylpyrimidin-2-yl)mercaptoacetyl]mutilin (I-30)

According to the synthetic method of Example 1, the compound I-30 of Example 30 was obtained by reacting intermediate E with 2-aminoethyl-1-ol.

HRMS(ES)calcd[M+H] ⁺ for C ₂₉ H ₄₃ N ₃ O ₅ S 546.3001, found 546.2999.

According to the synthesis method of Example 1, when intermediate C is 2-mercapto-5-methylpyrimidin-4-ol, intermediate E reacts with different small-molecule aliphatic amines to obtain compounds I-31～32 of Examples 31-32. I-32.

Example 31: Preparation of 14-O-[(4-(4-ethylpiperazin-1-yl)-5-methylpyrimidin-2-yl))mercaptoacetyl]mutilin (I-31)

According to the synthetic method of Example 1, the compound I-31 of Example 31 was obtained by reacting intermediate E with 1-ethylpiperazine.

mp: 81.3-83.6°C; ¹ H-NMR (400 MHz, CDCl ₃ ) δ 7.96 (d, J=5.9 Hz, 1H), 6.47 (dd, J=17.3, 11.0 Hz, 1H), 6.18 (d, J ＝6.1Hz,1H),5.73(d,J＝8.4Hz,1H),5.31(d,J＝11.0Hz,1H),5.17(d,J＝17.4Hz,1H),3.80(s,2H), 3.67(s, 4H), 3.34(s, 1H), 3.10(q, J=7.3Hz, 1H), 2.52(dd, J=15.9, 6.0Hz, 6H), 2.33–2.29(m, 1H), 2.19 (dq, J＝19.2, 8.6Hz, 3H), 2.08(s, 1H), 2.01(dd, J＝16.1, 8.5Hz, 2H), 1.75(d, J＝14.4Hz, 1H), 1.69–1.62( m, 2H), 1.56–1.45 (m, 3H), 1.40 (d, J=7.4Hz, 1H), 1.34 (d, J=16.3Hz, 2H), 1.27 (d, J=9.8Hz, 1H), 1.17-1.11 (m, 7H), 0.86 (d, J=6.8Hz, 3H), 0.72 (d, J=6.7Hz, 3H). ¹³ C-NMR (101MHz, CDCl ₃ )δ217.12, 168.36, 160.95, 155.75 ,139.12,117.09,98.88,69.52,58.19,52.31,52.20,45.45,44.56,43.94,43.57,41.89,36.79,35.98,34.49,34.14,30.44,26.89,26.42,24.82,16.76,14.93,11.72,11.45.HRMS (ES)calcd[M+H] ⁺ for C ₃₃ H ₅₀ N ₄ O ₄ S 599.3631, found 599.3634.

Example 32: Preparation of 14-O-[(4-((2-hydroxyethyl)amino)-5-methylpyrimidin-2-yl)mercaptoacetyl]mutilin (I-32)

According to the synthetic method of Example 1, the compound I-32 of Example 32 was obtained by reacting intermediate E with 2-aminoethyl-1-ol.

¹ H-NMR (400 MHz, CDCl ₃ ) δ 7.73 (s, 1H), 6.47 (dd, J=17.4, 11.0 Hz, 1H), 5.72 (d, J=8.5 Hz, 1H), 5.30 (d, J =11.0Hz,1H),5.17(d,J=17.7Hz,2H),3.82(dd,J=10.8,5.5Hz,3H),3.76(d,J=16.0Hz,1H),3.66(dt,J =10.6,5.2Hz,2H),3.34(d,J=6.1Hz,1H),2.34-2.26(m,2H),2.26-2.19(m,2H),2.19-2.12(m,1H),2.08( s, 1H), 2.04–1.99 (m, 1H), 1.97 (s, 3H), 1.75 (d, J=14.0Hz, 1H), 1.63 (dt, J=19.4, 10.5Hz, 3H), 1.56–1.46 (m,2H),1.43–1.29(m,3H),1.27(s,1H),1.13(s,4H),0.86(d,J=6.9Hz,3H),0.72(d,J=6.8Hz, 3H).HRMS(ES)calcd[M+H] ⁺ for C ₂₉ H ₄₃ N ₃ O ₅ S 546.3001, found 546.3001.

According to the synthesis method of Example 1, when intermediate C is 2-mercaptopyrimidin-4-ol, intermediate E reacts with different small-molecule fatty amines to obtain compounds I-33 to I-34 of Examples 33-34.

Example 33: Preparation of 14-O-[(4-(4-ethylpiperazin-1-yl)pyrimidin-2-yl)mercaptoacetyl]mutilin (I-33)

According to the synthetic method of Example 1, the compound I-33 of Example 33 was obtained by reacting intermediate E with 1-ethylpiperazine.

mp: 120.2-122.7°C; ¹ H-NMR (400 MHz, CDCl ₃ ) δ 7.96 (d, J=6.0 Hz, 1H), 6.47 (dd, J=17.4, 11.0 Hz, 1H), 6.18 (d, J ＝6.1Hz,1H),5.73(d,J＝8.4Hz,1H),5.31(d,J＝10.9Hz,1H),5.17(d,J＝17.4Hz,1H),3.80(s,2H), 3.67(s, 3H), 3.34(s, 1H), 3.11(q, J=7.3Hz, 1H), 2.52(d, J=9.8Hz, 4H), 2.48(d, J=7.1Hz, 1H), 2.33–2.29 (m, 1H), 2.19 (dq, J=19.2, 8.7Hz, 3H), 2.10–1.97 (m, 3H), 1.75 (d, J=14.2Hz, 1H), 1.67 (d, J= 12.1Hz, 1H), 1.63(s, 2H), 1.53(d, J=14.5Hz, 1H), 1.47(s, 1H), 1.41(s, 1H), 1.37(d, J=12.1Hz, 1H) ,1.32(s,1H),1.27(d,J＝9.8Hz,1H),1.19–1.10(m,7H),0.86(d,J＝6.8Hz,3H),0.72(d,J＝6.7Hz, 3H). ¹³ C-NMR (101MHz, CDCl ₃ )δ168.36,160.95,155.76,139.12,117.09,98.89,69.52,58.19,52.31,52.20,45.45,44.56,43.94,43.57,41.89,34.79,35. ,30.44,26.89,26.42,24.82,16.76,14.93,11.72,11.45.HRMS(ES)calcd[M+H] ⁺ forC ₃₂ H ₄₈ N ₄ O ₄ S 585.3474,found 585.3470.

Example 34: Preparation of 14-O-[(4-((2-hydroxyethyl)amino)pyrimidin-2-yl)mercaptoacetyl]mutilin (I-34)

According to the synthetic method of Example 1, the compound I-34 of Example 34 was obtained by reacting intermediate E with 2-aminoethyl-1-ol.

HRMS(ES)calcd[M+H] ⁺ for C ₂₈ H ₄₁ N ₃ O ₅ S 532.2845, found 532.2846.

According to the synthesis method of Example 1, when intermediate C is 2-mercapto-6-(trifluoromethyl)pyrimidin-4-ol, intermediate E is reacted with different small-molecule fatty amines to obtain the compounds of Examples 35-36 I-35 to I-36.

Example 35: 14-O-[(4-(4-ethylpiperazin-1-yl)-6-(trifluoromethyl)pyrimidin-2-yl)mercaptoacetyl]mutilin (I-35 ) preparation

According to the synthetic method of Example 1, the compound I-35 of Example 35 was obtained by reacting intermediate E with 1-ethylpiperazine.

mp: 164.2-166.3°C; ¹ H-NMR (400MHz, CDCl ₃ ) δ 6.51-6.42 (m, 2H), 5.71 (d, J=8.5Hz, 1H), 5.31 (d, J=11.0Hz, 1H) ), 5.17(d, J＝17.4Hz, 1H), 3.85(s, 2H), 3.73(s, 3H), 3.34(dd, J＝10.0, 6.7Hz, 1H), 2.54(d, J＝5.0Hz ,4H),2.51(s,1H),2.48(d,J=7.1Hz,1H),2.32–2.28(m,1H),2.24(dd,J=16.6,8.6Hz,2H),2.19–2.09( m, 1H), 2.07 (d, J=12.8Hz, 1H), 2.04–1.99 (m, 1H), 1.75 (d, J=13.6Hz, 1H), 1.67 (d, J=12.6Hz, 1H), 1.62(d,J＝10.6Hz,1H),1.55(d,J＝13.7Hz,1H),1.47(d,J＝9.9Hz,3H),1.40(d,J＝7.5Hz,1H),1.36( s,1H),1.32(s,1H),1.27(d,J=8.6Hz,1H),1.16(s,1H),1.13(d,J=4.3Hz,6H),0.86(d,J=6.9 Hz, 3H), 0.71 (d, J=6.9 Hz, 3H). ¹³ C-NMR (101 MHz, CDCl ₃ ) δ 217.06, 167.94, 161.29, 138.99, 117.11, 95.14, 69.83, 58.13, 52.25, 52.13, 45.44, 44.55 ,43.97,41.90,36.75,35.95,34.48,34.12,30.42,26.86,26.25,24.80,16.78,14.85,11.76,11.44.HRMS(ES)calcd[M+H] ⁺ for C ₃₃ H ₄₇ F ₃ N ₄ O 4S _653.3348 , found 653.3351.

Example 36: Determination of 14-O-[(4-((2-hydroxyethyl)amino)-6-(trifluoromethyl)pyrimidin-2-yl)mercaptoacetyl]mutilin (I-36) preparation

According to the synthetic method of Example 1, the compound I-36 of Example 36 was obtained by reacting intermediate E with 2-aminoethyl-1-ol.

HRMS(ES)calcd[M+H] ⁺ for C ₂₉ H ₄₀ F ₃ N ₃ O ₅ S 600.2719, found 600.2715.

According to the synthesis method of Example 1, intermediate B and intermediate C-2 generate intermediate D, intermediate D undergoes a sulfonylation reaction, and intermediate E reacts with different small-molecule aliphatic amines to obtain the compounds of Examples 37-38. Compounds 1-37 to 1-38.

Example 37: 14-O-[(4-(4-Ethylpiperazin-1-yl)-5-methylthieno[2,3-d]pyrimidin-2-yl)mercaptoacetyl]mer Preparation of Lin (I-37)

According to the synthetic method of Example 1, intermediate E was reacted with 1-ethylpiperazine to obtain compound I-37 of Example 37.

mp: 108.6-109.3°C; ¹ H-NMR (400 MHz, CDCl ₃ ) δ 6.81 (s, 1H), 6.46 (dd, J=17.4, 11.1 Hz, 1H), 5.72 (d, J=8.5 Hz, 1H) ),5.28(d,J＝10.9Hz,1H),5.14(d,J＝17.4Hz,1H),3.91(d,J＝16.3Hz,2H),3.58(s,4H),3.33(s,1H ), 2.69(s, 4H), 2.57(d, J=8.3Hz, 3H), 2.48(s, 3H), 2.30(d, J=6.9Hz, 1H), 2.24–2.19(m, 2H), 2.07 (s,1H),2.00(dd,J=16.1,8.5Hz,2H),1.75(d,J=13.0Hz,2H),1.63(d,J=10.2Hz,2H),1.53(d,J= 14.2Hz, 1H), 1.37(s, 1H), 1.32(s, 1H), 1.27(d, J=10.1Hz, 2H), 1.18(t, J=7.2Hz, 3H), 1.12(s, 3H) , 1.09(s, 1H), 0.85(d, J=6.9Hz, 3H), 0.76(d, J=6.8Hz, 3H). ¹³ C-NMR(101MHz, CDCl ₃ )δ217.12,168.23,162.20,138.95, 129.36,118.13,117.17,69.64,58.15,52.42,52.01,49.80,45.45,44.60,43.97,41.90,36.82,35.96,34.49,34.34,30.44,26.89,26.28,24.82,17.01,16.97,14.91,11.51,11.45. HRMS(ES)calcd[M+H] ⁺ for C ₃₅ H ₅₀ N ₄ O ₄ S ₂ 655.3351, found 655.3353.

Example 38: 14-O-[(4-((2-hydroxyethyl)amino)-5-methylthieno[2,3-d]pyrimidin-2-yl)mercaptoacetyl]mutilin ( Preparation of I-38)

According to the synthetic method of Example 1, the compound I-38 of Example 38 was obtained by reacting intermediate E with 2-aminoethyl-1-ol.

HRMS(ES)calcd[M+H] ⁺ for C ₃₁ H ₄₃ N ₃ O ₅ S ₂ 602.2722, found 602.2721.

Effect Example

In vitro antibacterial experiments of compounds I-1 to I-38 provided by the present invention

Determination of Minimum Inhibitory Concentration (MIC)

The experiment used the micro-broth dilution method. The experimental control drugs were tiamulin and ritamulin. Tiamulin, a pleuromutilin antibiotic, is one of the top ten veterinary antibiotics in the world. Retamorelin, as the first human pleuromutilin antibiotic, was approved by the US FDA in 2007. All synthetic pleuromutilin derivatives are directed against S. aureu s, S. agalactiae, E. coli and some synthetic pleuromutilin derivatives Determination of MIC against Methicillin-Resistant Staphylococcus aureus (MRSA)

Preparation of target compound stock solution: accurately weigh 6.4 mg of target compound and place it in a 10 mL volumetric flask, dissolve with 0.25 mL of DMSO, add 9.5 mL of distilled water, 0.25 mL of Tween 80 to volume, and shake well to obtain a stock solution (6.4 mg /mL), sterilized with a 0.22 μm filter, aliquoted into small tubes, and stored at -20°C. The control drugs Tiamulin and Retamorelin were also prepared according to the above method.

Preparation of bacterial liquid: Take out the well-preserved strains at -20 °C and inoculate them on a new MH plate. After culturing at 37 °C for 24 hours, pick a single colony and inoculate it in MH medium for another 24 hours; select a single colony and transfer it to a sterile physiological The turbidity was adjusted to 0.6McF in saline, and the bacterial concentration was 10 ⁶ CFU/mL at this time.

MIC plate preparation: Dilute the target compound stock solution (6.4 mg/mL) by 10 times to obtain the target compound solution with a concentration of 640 μg/mL; take a sterile 96-well plate, add 180 μL MH broth medium to the first hole, and add 180 μL MH broth to the first hole. Add 100 μL of MH broth medium to wells 2 to 10 respectively, add 20 μL of antibacterial drugs at a concentration of 640 μg/mL to the first well, and add 100 μL to the second well after mixing. By analogy, the 12th well draws 100 μL and discards it. Finally, 100 μL of the diluted bacterial solution was added to each well, and three replicates were performed for each bacterial strain. At this time, the drug concentrations in each well were: 64, 32, 16, 8, 4, 2, 1, 0.5, 0.25, 0.125, 0.06, 0.03, 0.015 μg/mL, and three groups of drugs at each concentration were made in parallel.

Inoculated bacterial solution: add 100 μL bacterial solution to each well 1 to 12, so that the final bacterial solution concentration of each well is about 5×10 ⁵ CFU/mL, and the drug concentration of well 1 to well 12 is 32, 16, 8, and 4, respectively. , 2, 1, 0.5, 0.25, 0.125, 0.06, 0.03, 0.015 μg/mL. The inoculated 96-well plate was placed in a 37°C incubator for cultivation, and the growth of bacterial liquid was observed for 24 hours. The control drugs tiamulin and ritamulin are determined by the same method, and the lowest drug concentration that completely inhibits the growth of bacteria in the small well is the MIC, and the bacteria in the positive control well (ie, no drug) need to grow significantly. When there is a single jump hole in the micro-broth dilution method, record the highest drug concentration that inhibits bacteria, and repeat the test if there are multiple jump holes.

Table 1 In vitro minimum inhibitory concentration data

It can be seen from Table 1 that the pleuromutilin derivatives with pyrimidine side chains provided by the present invention have strong antibacterial activity.

The target compounds I-1~I-38 basically showed good antibacterial activity against Staphylococcus aureus and Streptococcus agalactiae, and compounds I-1 and I-16 were resistant to Staphylococcus aureus, Streptococcus agalactiae and methoxy-resistance. The antibacterial activity of Staphylococcus aureus is better than that of Tiamulin and Retamorelin, which are widely used in clinical practice.

Time-kill curve determination

According to the minimum inhibitory concentration data in Table 1, the compounds I-1 and I-16 with the best effect, that is, the smaller values of the minimum inhibitory concentration, were selected for in vitro bactericidal curve experiments. S. aureus was cultured in LB broth, diluted to 10 ⁶ CFU/mL, and compounds I-1 and I-16 were added at concentrations of 1×MIC, 4×MIC, 8×MIC, 16×MIC, and 32×MIC, respectively. Co-culture with Tiamulin liquid, take 100 μL of bacterial liquid at 0, 3, 6, 9, 12 and 24 h, and serially dilute it with sterile normal saline 10 times. Plate 100 µL of the dilution on LB agar plates. After culturing at 37 °C for 20 h, the total number of colonies CFU/mL was calculated using Image J software. Taking time as the abscissa and log10CFU/mL of the total number of bacteria as the ordinate, the time-killing curve was drawn using GraphPad Prism software.

The results are shown in Fig. 1, which shows the time-killing curves of different concentrations of compounds I-1, I-16 and Tiamulin on S. aureus. It can be seen from the figure that compounds I-1 and I-16 can completely eliminate S. aureus at 1×MIC, which are time-dependent bactericidal agents.

Claims (10)

1. A pleuromutilin derivative with a pyrimidine side chain is a compound shown as a formula I or a stereoisomer, a pharmaceutically acceptable salt or a crystal form thereof:

wherein,

R ₁ 、R ₂ each independently selected from H and (C) ₁ -C ₆ ) Alkyl, 1-3 halogen substituted (C) ₁ -C ₆ ) An alkyl group; or when R is ₁ And R ₂ When the position has no substituent, R ₁ And R ₂ Are linked to other atoms to form a cyclic structure A selected from (C) ₆ -C ₁₀ ) Aryl, 5-10 membered heteroaryl, 5-10 membered heterocyclyl, said heteroaryl and heterocyclyl containing 1-3 heteroatoms selected from N, O, S and being interrupted by 0-3R which may be the same or different _A Substitution;

R _A selected from H, halogen, cyano, nitro, hydroxyl, carboxyl, amino, (C) ₁ -C ₆ ) Alkyl, 1-3 halogen or amino or hydroxy or cyano substituted (C) ₁ -C ₆ ) Alkyl, (C) ₁ -C ₆ ) Alkylamino radical, di (C) ₁ -C ₆ ) Alkylamino radical, (C) ₁ -C ₆ ) Alkoxy group, (C) ₁ -C ₆ ) Alkylcarbonyl group, (C) ₂ -C ₆ ) Alkenyl, halo (C) ₂ -C ₆ ) An alkenyl group;

R ₃ selected from H, fatty amino,

B is selected from (C) ₆ -C ₁₀ ) Aryl, 5-10 membered heteroaryl, and substituted with 0-3R which may be the same or different _B Substitution;

R _B selected from H, halogen, cyano, nitro, hydroxyl, carboxyl, amino, (C) ₁ -C ₆ ) Alkyl, 1-3 halogen or amino or hydroxy or cyano substituted (C) ₁ -C ₆ ) Alkyl, (C) ₁ -C ₆ ) Alkylamino radical, di (C) ₁ -C ₆ ) Alkylamino radical, (C) ₁ -C ₆ ) Alkoxy, 1-3 halogen or amino or hydroxy or cyano substituted (C) ₁ -C ₆ ) Alkoxy group, (C) ₁ -C ₆ ) Alkylthio group, (C) ₁ -C ₆ ) Alkylcarbonyl group, (C) ₁ -C ₆ ) Alkylsulfonyl group, (C) ₂ -C ₆ ) Alkenyl, halo (C) ₂ -C ₆ ) An alkenyl group.

2. A pleuromutilin derivative with a pyrimidine side chain as claimed in claim 1 wherein in formula I:

R ₁ selected from H, (C) ₁ -C ₆ ) Alkyl, methyl substituted with 1-3 fluorines;

R ₂ selected from H, (C) ₁ -C ₆ ) An alkyl group;

or when R is ₁ And R ₂ When the position has no substituent and is connected with other atoms to form a cyclic structure A, A is selected from (C) ₆ -C ₁₀ ) Aryl, 4-6 membered heteroaryl, 4-6 membered heterocyclyl, said heteroaryl and heterocyclyl containing 1-3 heteroatoms selected from N, O, S and being interrupted by 0-3R which may be the same or different _A Substitution;

R _A selected from H, halogen, amino, (C) ₁ -C ₆ ) Alkyl, 1-3 halogen or amino or hydroxy or cyano substituted (C) ₁ -C ₆ ) Alkyl, (C) ₁ -C ₆ ) Alkoxy group, (C) ₁ -C ₆ ) Alkylcarbonyl group, (C) ₂ -C ₆ ) An alkenyl group;

R ₃ selected from H,

R _3a And R _3b The same or different are respectively and independently selected from H and (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl, (C) ₂ -C ₆ ) Alkynyl, halogen or amino or hydroxy or cyano substituted (C) ₁ -C ₆ ) An alkyl group; or R _3a And R _3b Together with the nitrogen atom to which they are attached form a 4-10 membered heterocyclic group, except for R _3a And R _3b Optionally containing, in addition to the nitrogen atom to which it is attached, 1 to 4 heteroatoms selected from N, O, S or sulfone or carbonyl groups, said heterocyclic group optionally being substituted with 0 to 3R, which may be the same or different _3c Substitution;

R _3c selected from H, halogen, cyano, nitro, hydroxyl, carboxyl, amino, (C) ₁ -C ₆ ) Alkyl, 1-3 halogen or amino or hydroxy or cyano substituted (C) ₁ -C ₆ ) Alkyl, (C) ₁ -C ₆ ) Alkylamino radical, di (C) ₁ -C ₆ ) Alkylamino radical, (C) ₁ -C ₆ ) Alkoxy group, (C) ₁ -C ₆ ) Alkylcarbonyl group, (C) ₁ -C ₆ ) Alkylsulfonyl group, (C) ₁ -C ₆ ) Alkylsulfonylamino group, (C) ₁ -C ₆ ) Alkylamido radical, (C) ₂ -C ₆ ) Alkenyl, halo (C) ₂ -C ₆ ) Alkenyl, alkylamino substituted (C) ₂ -C ₆ ) Alkenyl, (C) ₃ -C ₇ ) A cycloalkyl group;

b is selected from (C) ₆ -C ₁₀ ) Aryl, 5-membered heteroaryl, and substituted by 0-2R which may be the same or different _B Substitution;

R _B selected from H, halogen, nitro, hydroxy, (C) ₁ -C ₆ ) Alkyl, 1-3 halogen substituted (C) ₁ -C ₆ ) Alkyl, (C) ₁ -C ₆ ) Alkoxy group, (C) ₁ -C ₆ ) Alkylthio group, (C) ₁ -C ₆ ) An alkylcarbonyl group.

3. A pleuromutilin derivative with a pyrimidine side chain as claimed in claim 2 wherein in formula I:

R ₁ selected from H, methyl, trifluoromethyl;

R ₂ selected from H, methyl;

or when R is ₁ And R ₂ When the position has no substituent and is connected with other atoms to form a cyclic structure A, A is selected from (C) ₆ -C ₁₀ ) Aryl, 4-6 membered heteroaryl, containing 1-3 heteroatoms selected from N, O, S, and substituted with 0-3R, the same or different _A Substitution;

R _A selected from H, methyl;

R ₃ selected from H,

R _3a And R _3b The same or different, are respectively and independently selected from H and (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl, (C) ₂ -C ₆ ) Alkynyl, halogen or amino or hydroxy or cyano substituted (C) ₁ -C ₆ ) An alkyl group; or R _3a And R _3b Together with the nitrogen atom to which they are attached form a 4-10 membered heterocyclic group, except for R _3a And R _3b Optionally containing, in addition to the nitrogen atom to which it is attached, 1 to 4 heteroatoms selected from N, O, S or sulfone or carbonyl groups, said heterocyclic group optionally being substituted with 0 to 3R, which may be the same or different _3c Substitution;

R _3c selected from H, halogen, cyano, nitro, hydroxyl, carboxyl, amino, (C) ₁ -C ₆ ) Alkyl, 1-3 halogen or amino or hydroxy or cyano substituted (C) ₁ -C ₆ ) Alkyl, (C) ₁ -C ₆ ) Alkylamino radical, di (C) ₁ -C ₆ ) Alkylamino radical, (C) ₁ -C ₆ ) Alkoxy group, (C) ₁ -C ₆ ) Alkylcarbonyl group, (C) ₁ -C ₆ ) Alkylsulfonyl group, (C) ₁ -C ₆ ) Alkylsulfonylamino group, (C) ₁ -C ₆ ) Alkylamido radical, (C) ₂ -C ₆ ) Alkenyl, halo (C) ₂ -C ₆ ) Alkenyl, alkylamino substituted (C) ₂ -C ₆ ) Alkenyl, (C) ₃ -C ₇ ) A cycloalkyl group;

b is selected from (C) ₆ -C ₁₀ ) Aryl, 5-membered heteroaryl, and substituted by 0-2R which may be the same or different _B Substitution;

R _B selected from H, halogen, nitro, hydroxy, (C) ₁ -C ₆ ) Alkyl, 1-3 halogen substituted (C) ₁ -C ₆ ) Alkyl, (C) ₁ -C ₆ ) Alkoxy group, (C) ₁ -C ₆ ) Alkylthio group, (C) ₁ -C ₆ ) An alkylcarbonyl group.

4. A pleuromutilin derivative with a pyrimidine side chain as claimed in claim 3, wherein in formula I:

R ₃ is selected from

R _3a And R _3b The structure consisting of the nitrogen atom to which it is attached is as follows:

b is selected from phenyl, thienyl, and substituted by 0-2R which may be the same or different _B Substitution;

R _B selected from H, nitro, methyl, trifluoromethyl, methoxy and halogen.

5. The pleuromutilin derivative with a pyrimidine side chain as claimed in claim 4, wherein said pleuromutilin derivative with a pyrimidine side chain is any one of compounds I-1 to I-38 as shown below:

6. a pharmaceutical composition comprising a pleuromutilin derivative having a pyrimidine side chain as claimed in any one of claims 1 to 5 and a pharmaceutically acceptable carrier, excipient, diluent, adjuvant, vehicle or combination thereof.

7. Use of a pleuromutilin derivative having a pyrimidine side chain as claimed in any one of claims 1 to 5 in the manufacture of an antibacterial medicament.

8. Use of the pharmaceutical composition of claim 6 for the preparation of an antibacterial medicament.

9. Use of a pleuromutilin derivative having a pyrimidine side chain as claimed in any one of claims 1 to 5 or a pharmaceutical composition as claimed in claim 6 in the manufacture of a medicament for the treatment of staphylococcus aureus, methicillin-resistant staphylococcus aureus or streptococcus agalactiae.

10. A process for the preparation of a pleuromutilin derivative having a pyrimidine side chain as claimed in claim 5, comprising the steps of:

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