(+)-Thienamycin

Thienamycin is a member of carbapenems.

Method 1: Fermentation method. 1. Seed culture (1) Into a 250ml conical flask equipped with a baffle, transfer 50ml of sterile solvent A, and under aseptic conditions, suspend a test tube of freeze-dried Streptomyces cattleya strain in the solution. The composition of solvent A is as follows: autolyzed yeast 10.0g, glucose 10.0g, phosphate buffer 2.0ml, MgSO4-7H2O0.05g, distilled water 1000ml, and adjust the pH value to 6.5 with sodium hydroxide. (2) The culture flask was shaken (220r/min) at 28°C for 48h. Under aseptic conditions, remove 40 ml of the seed culture solution and mix with 40 ml of sterile 20% (V/V) ethylene glycol. Transfer 2ml of this mixed solution to a sterile vial with a pipette, freeze and store in liquid nitrogen. (3) Transfer 50ml of solvent A into a 250ml conical flask equipped with a baffle, and then implant the contents of the above freezing vial. The seed bottle was shaken (160r/min) at 28°C for 24h. (4) Transfer 500ml of solvent A into a 2L Erlenmeyer flask equipped with a baffle, and implant 10ml of the culture solution from the upper seed bottle. Incubate for 24h with shaking (160r/min) at 28°C. (5) Transfer 67L of solvent A into a 756L stainless steel fermentor, and plant 1000ml of the seed liquid obtained in (4). Stirring (130r/in) and aerating (0.283rn3/min) at 28°C for 24h. if needed. Polyethylene glycol 2000 with no more than 0.1% can be added as a defoaming agent. 2. Fermentation Transfer 4082L solvent E into the 5670L stainless steel fermentor, and implant 453L (5) of the liquid obtained. Stirring (70r/min) and air filling (54.3cfm) at 24°C for 144h. If necessary, no more than 0.1% polyethylene glycol 2000 can be added as a defoaming agent. The composition of solvent E is as follows: 25.0g of cerelose, 15.0g of corn extract (wet), 10.0g of distiller’s grains, 5.0g of medicinal cottonseed liquid, 0.01g of CoCl2?6H2O, 3.0g of CaCO3 (after pH adjustment) ), polyethylene glycol 2000 0.25%, tap water 1000ml; adjust the pH to 7.3 with sodium hydroxide. 3. Separation and extraction (1) Filter the 4082L fermentation broth obtained with 4% (W/V) filter aid. Add 12g of sodium salt of ethylenediaminetetraacetic acid to the filtrate and cool to 6°C. Keep it at 613 and adjust the pH to 4.5±0.2. At a rate of 48L/min, 480L of 20-50μm Dowex 50×4 Na+ was used to adsorb the cold filtrate. Wash the adsorbent with 480L deionized water, and then wash it with 2% pyridine aqueous solution at a rate of 24L/min. Collect 3 eluates, each of 300L, 520L and 240L, each containing 4%, 16% and 6% of the active substance. The second eluate was concentrated to 48L and adjusted to pH=7. (2) The 48L concentrated solution was adjusted to pH=7.3, and was adsorbed with 76L 50-100μm Dowex 1×2, and the flow rate was 7.6L/min. Elute with deionized water at the same speed. Collect 4 eluates, each of 48L, 48L, 70L and 48L, and adjust the pH to 7. It was identified that the 70L eluate contained 68% of the active substance used for adsorption. The eluate was concentrated to 18L, adjusted to pH 7.0, and lyophilized to obtain 99g product, 310U/mg. (3) 10 g of the freeze-dried solid is dissolved in a 0.1 mol/L 2, 6-lutidine acetate buffer (PH=6.3). Then adjust the pH to 6.3 with acetic acid, and perform chromatography on a 7.6 X 142 cm Dowex 50 X 8 (200-400 μm) column. The eluent is 0.1 mol/L buffer, and the elution rate is 25 ml/min. Collect the first 3L, 200 20ml effluent. Combine the effluent from the 80th to the 136th part and dilute with 590ml deionized water to obtain 1760ml liquid. This fluid contains 62% of the active substance used for chromatography. (4) The 1760ml liquid obtained above is freeze-dried. The lyophilized solid was dissolved in about 27 ml of 0.1 mol/L 2, 6-lutidine acetate (pH=7.0). Use Bio-Gel P-2 (200～400μm) 5×112cm column for chromatography, the eluent is the same buffer, and the speed is 10ml/min. Collect 105 effluents of 20 ml each. After identification, the 75th to 80th effluents were combined. After lyophilization, 90mg thiomycin was obtained, 10000U/mg. Method 2: The synthetic method can use penicillin as raw materials, see KaradyS, amato J S, Reamer RA and Weinstock L M. Stereospecific conversion of Penicillin to Thienamycin. J AC S, 1981, 103(22): 6765-6767; Salzmann T, Ratcliffe RW, ChristensenB Gand Bouffard FA. A stereocontrolled synthesisof(+)-Thienamycin. J AC S, 1980, 102(19): 6161-6163; Kametani T, Huang SP, Nakayama A and Hon. Da T. Further studies on the synthesis of Thienamycin: a facile and stereoseleetive synthesis of a bicyclic β-keto ester by 1, 3-dipolar cycloaddition. J Org Chin, 1982, 47(12): 2328--2331.

The first natural carbapenem antibiotic has the characteristics of broad spectrum, high efficiency, and stability to bacterial β-lactamase. However, the chemical nature is unstable and it is difficult to apply it directly to the clinic.

Molecular Weight:272.32
XLogP3:-3
Hydrogen Bond Donor Count:3
Hydrogen Bond Acceptor Count:6
Rotatable Bond Count:5
Exact Mass:272.08307817
Monoisotopic Mass:272.08307817
Topological Polar Surface Area:129
Heavy Atom Count:18
Complexity:423
Defined Atom Stereocenter Count:3
Covalently-Bonded Unit Count:1
Compound Is Canonicalized:Yes