133results about "Polymyxins" patented technology

Novel protected cyclopeptide intermediates are prepared from polymyxin B are used to synthesize new peptide antibiotics. Intermediates are readily derivatized and deprotected to provide new families of antibiotics, which have potent anti-bacterial activity against gram-negative bacteria; but also are useful and potent against gram-positive bacteria.

Novel conjugates of bacterial outer membrane binding peptides, preferably having bacterial sensitization activity, and immune cells chemotactic peptides, and pharmaceutical compositions containing same useful in the treatment of bacteremia and / or septicemia following infection by gram negative bacteria administered alone or in combination with conventional antibiotics.

The present invention provides a new class of polymyxin derivates useful for treating bacterial infections, especially Gram-negative infections, that have reduced renal cytotoxicity.

Provided herein are novel compounds and novel protected compounds that can be derived from polymyxin, including, e.g., polymyxin A. The novel compounds have antibacterial properties against a diverse range of Gram negative bacteria and reduced toxicity compared to polymyxins such as polymyxin A. Also provided are antibacterial pharmaceutical compositions containing the novel compounds and novel protected compounds, as well as methods for preparing the antibacterial compounds and protected compounds.

Provided herein are novel compounds and novel protected compounds that can be derived from polymyxin, including, e.g., polymyxin B1 and [Il7] polymyxin B1. The novel compounds have antibacterial properties against a diverse range of gram negative bacteria and reduced toxicity compared to polymyxins such as polymyxin B. Also provided are antibacterial pharmaceutical compositions containing the novel compounds and novel protected compounds, as well as methods for preparing the antibacterial compounds and protected compounds.

The invention discloses a preparing method of high-purity polymyxin B. The preparing method comprises the steps of preparation of column-loading solution; injecting the column-loading solution into a high performance liquid chromatography preparative column, wherein sodium sulfate - phosphate buffer / organic solvent is used as mobile phase, collecting target components, removing impurities, especially B1 - 1 and B3, and determining collected solution by means of high performance liquid chromatography (HPLC), wherein chromatographic purity of the target components B1 or B2 or mixture of the B1 and the B2 is more than 99.5%; and purification, wherein vacuum concentration is carried out on the collected target components so that the organic solvent is removed, organic solvent desalination concentration is carried out on balance phase, concentrated solution is spray-dried or freeze-dried to produce the polymyxin B with purity of more than 99.5%. The preparing method of the high-purity polymyxin B has the advantages of being high in purity, high in yield, simple in process, and capable of injecting sample solution continuously. Quality of products is far better than the medical standard regulated by the European Pharmacopoeia (EP) and the United States Pharmacopeia (USP). One-time investment of equipment is slightly high, while operation cost is not high, and therefore the preparing method of the high-purity polymyxin B is suitable for industrial production.

The invention relates to a method for extracting polymyxin B and E from a fermentation technique culture, which comprises the following steps of: adjusting the pH value of the fermentation technique culture to 10.0 to 14.0 by utilizing an alkaline substance under the temperature of 0 to 25 DEG C and then stirring for 30 min; collecting fungus dregs by utilizing a solid-liquid separation technique to obtain a polymyxin B and E free alkali precipitate; adjusting the pH value of the free alkali precipitate obtained by the solid-liquid separation technique to 1.0 to 6.0 by using an acid substance or solution; after the polymyxin B and E free alkali precipitate is dissolved, carrying out solid-liquid separation to obtain the concentrated solution of polymyxin B and E salts; removing inorganic salts by using a nanofiltration technology; and finally carrying out spray drying to obtain the finished product. The method is characterized by extracting the polymyxin B and E from fermentation liquor in a convenient and high-efficiency mode, solves the problem of high production cost because of high labor strength, long working procedure and low extraction yield in the production process of other extraction methods, and has the advantages of no use of solvents, low cost, environment protection, high yield and low characteristic requirements on the fermentation liquor and equipment.

The invention discloses a process for extracting colistin sulfate through a precipitation method. The process comprises the following steps of: (a) adjusting pH of fermentation liquor containing polymyxin E with oxalic acid or concentrated sulfuric acid, and filtering with a filtering device; (b) adding 0 metal complexing agent and inorganic flocculant to the filtrate, and reacting at normal temperature for 0.5 to 4.0h; (c) heating, cooling, and vacuum-filtering the reactive liquid; (d) adjusting pH of the filtrate above with aqueous alkali, separating the solid from the liquid to obtain a filter cake, dissolving the filter cake with sulfuric acid solution so as to obtain acidizing fluid, adding potassium permanganate and activated carbon to the acidizing fluid respectively, agitating for0.5 to 2.0 hours, and then vacuum-filtering to obtain colistin sulfate liquid; and (e) centrifugally spray-drying the colistin sulfate liquid. With the adoption of the process, the common problems ofthe existing process for purifying fermentation liquor that the steps are complicated, the period is long, the labor intensity is relatively high, and the wastewater is difficult to treat and the like can be overcame.

The present invention provides a method for separation purification of polymyxin B1 from a polymyxin B mixed component. The method comprises: (a) loading a chromatography medium of a macroporous absorption resin to obtain a chromatography column; and (b) adding a polymyxin B solution, and adopting an organic solvent-containing acid solution as an eluent to sequentially elute various components of the polymyxin B so as to obtain the polymyxin B1 solution after elution. According to the method, the operation is simple, the adopted separation medium has characteristics of stable chemical characteristic and high re-use rate, industrial scale-up is easily achieved, and the peak purity of the obtained polymyxin B1 can be more than 98%.

The invention discloses a method for extracting polymyxin B from a fermentation broth of the polymyxin B. The method for extracting the polymyxin B from the fermentation broth of the polymyxin B comprises the following steps: a, conducting solid-liquid separation on the fermentation broth of the polymyxin B and obtaining a filtrate; b, extracting the filtrate with a polar organic solvent, and extracting an extract liquor reversely with water when the extract liquor is at the potential hydrogen (pH) value of 1.0-2.0; c, adding the reversal extract liquor which is obtained from the step b into inorganic salt, wherein the volume of the reversed extract liquor of the inorganic salt is 3-8% (w / v), and adjusting the pH value to be 9.0-10.0 so that the polymyxin B is obtained after precipitation and sediment. Preferably, the method for extracting the polymyxin B from the fermentation broth further comprises a step of d: reacting the polymyxin B and sulfuric acid to form salt, and dissolving and precipitating, thereby obtaining the polymyxin B sulfate. The polymyxin B or the sulfate prepared by means of the method is higher in yield and purity, the purity of a product is above 85%, the tilter is above 8100IU / g, and the yield is above 60%. The method for extracting polymyxin B from the fermentation broth has the advantages of being easy and convenient to operate, short in time consuming, low in requirement of equipment, and capable of recycling the organic solvent and being used for large-scale industrial production.

The present invention provides, in part, a Paenibacillus sp. isolate, designated Paenibacillus polymyxa JB05-01-1, as well as an anti-microbial agent obtained from the bacterium or cell culture supernatant thereof. Compositions, methods and uses are also provided.

The invention belongs to the technical field of medicine, and particularly relates to a polymyxin E2 composition, and a preparation method and application thereof. The composition mainly contains polymyxin E2, and also comprises E1, E3, E1-1, E1-7M0A and the like. The application provided by the invention is the application of the composition in preparation of medicaments for treating gram-negative bacterial infection.

The present invention relates to a polymyxin derivative wherein R1, R2 and R3 are optional and R1, R2, R3, R5, R8 and R9 are cationic or neutral amino acid residues selected so that the total number of positive charges at physiological pH is at least two but no more than three; and to a combination product comprising at least two such derivatives. The invention further relates to a method for treating, alleviating or ameliorating an infection in a subject, caused by a Gram-negative bacterium by administering a therapeutically effective amount of a derivative according to the present invention to said subject; to a method for sensitizing Gram-negative bacteria to an antibacterial agent by administering, simultaneously or sequentially in any order a therapeutically effective amount of said antibacterial agent and a derivative according to the present invention to said subject; to methods for developing novel antibiotics; for reducing the nephrotoxicity, for improving the pharmacokinetic properties of natural polymyxins and octapeptins; and for sensitizing clinically important bacteria to a host defence mechanism complement present in serum. Finally, the invention relates to a process for preparing such polymyxin derivatives.

Provided are a composite carrier for immobilization of a protein, polypeptide or oligopeptide, a preparation method and an application thereof. The composite carrier is a porous material which comprises: (1) a porous organic foam material containing open pores; and (2) a crosslinked product having aldehyde groups and immobilized on the surface of the walls of one or more pores of the porous organic foam material, wherein the aldehyde groups are able to react with the protein, polypeptide or oligopeptide, and the crosslinked product are formed by chitosan via a crosslinking reaction with a polyaldehyde compound.

The invention provides a culture medium for fermenting polymyxin B sulfate. Each 100ml of the culture medium is prepared from the following components: 6g to 12g of flour, 0.03g to 0.06g of high-temperature amylase, 0.4g to 2g of cottonseed meal, 0.3g to 1g of corn pulp, 0.01g to 0.05g of dipotassium phosphate, 0.2g to 0.6g of ammonium sulfate, 0.2g to 0.6g of calcium carbonate, 0.02ml to 0.08ml of a de-foaming agent and the balance of water. A method for fermenting and producing the polymyxin B sulfate comprises the following the steps: inoculating the culture medium with strain seed liquid for producing the polymyxin B sulfate; fermenting and culturing fermentation liquid at 26 DEG C to 32 DEG C, wherein the pH (Potential of Hydrogen) value is 6.7 to 7.4 at the beginning of fermenting; controlling the pH value of the fermentation liquid to be natural during a fermentation process, wherein the dissolved oxygen volume concentration of the fermentation liquid is 10 percent to 30 percent. All the components of the culture medium are cooperatively matched and the content of impurities B2P is controlled to be lower than 1.5 percent.

The invention discloses a method for extraction of colistin sulphate. The method comprises the following steps of 1, a fermentation broth preparation process comprising adjusting a pH value to 4.0 by sulfuric acid, 2, a ceramic membrane filtration process comprising filtering the fermentation broth by a ceramic membrane, collecting a filtrate, and adjusting a pH value of the filtrate to 4.5-5, 3, a continuous moving bed absorption-desorption process, wherein through the continuous moving bed absorption-desorption process, a discharged liquid is obtained, 4, a discharged liquid decoloration process, 5, a nanofiltration concentration process, wherein in the nanofiltration concentration process, nanofiltration membrane pore sizes are in a range of 2100-400Da molecular weight cut-off and 10-20 times concentration is realized, and 6 a spray drying process. The method has a total colistin sulphate yield of 86 to 91% and has good energy-saving and emission-reduction effects.

The invention provides a method for producing polymyxin E through fermentation and foam separation coupling. According to the method, polymyxin E is prepared by a circulating reflux device formed by a peristaltic pump through fermentation, film separation and foam separation coupling. During coupling operation, fermentation liquor enters a foam separation tower through a film filter by means of the peristaltic pump, products and a liquid pass through the film while cells are intercepted in a fermentation tank, fermentation continues to be performed, and constancy of bacteria is guaranteed; a foam distributor is mounted at the bottom of the foam separation tower, bubbles are produced through air blowing, foam separation is performed, part of nutrients enter the foam separation tower through the film filter, a remaining liquid produced after foam separation returns to the fermentation tank in a circulating reflux manner, and sufficient utilization of the nutrients is guaranteed; coupling outside a tank body is adopted, operation conditions for fermentation and operation conditions for foam separation are controlled respectively, and the best operation conditions can be met.

The invention discloses a preparation method of colistin sulfate premixing agents. The PH of the colistin sulfate fermentation liquid is regulated to 4.5 to 5.0 by sulfuric acid, the plate frame filtering and the re-filtering are carried out to obtain wet germ slag and filter liquid, the filter liquid is concentrated by a dual-effect concentrator, and the concentration liquid is obtained; the concentration liquid and the wet germ slag are mixed into mixed liquid, ground limestone and defatted rice bran are added into the mixed liquid, the materials are uniformly mixed, and a mixture is obtained; and the mixture is subjected to expansion drying, the air inlet temperature of expansion drying agents is controlled to be 125 to 135 DEG C, the air outlet temperature is 60 to 70 DEG C, and finished products of the colistin sulfate premixing agents are obtained. The preparation method provided by the invention has the advantages that the utilization rate is high, the yield is high, and the operation is easy.

The invention provides a sulfate polymyxin B crystal and a preparation method thereof. The preparation method comprises the following steps: firstly, adsorbing polymyxin B fermented liquid with resin,and adding a sulfate aqueous solution and / or an acid ethanol aqueous solution for desorption to obtain a desorption solution; secondly, adjusting a pH value of the desorption solution obtained in thefirst step to 5.0 to 7.0; and then concentrating until a solid is precipitated to obtain a sulfate polymyxin B saturated solution; and thirdly, in a stirring state, dropwise adding an organic solventinto the sulfate polymyxin B saturated solution obtained in the second step or dropwise adding the sulfate polymyxin B saturated solution obtained in the second step into the organic solvent so as toseparate out the crystal, filtering and drying a filter cake, thus obtaining the sulfate polymyxin B crystal. The solvent used in the preparation method disclosed by the invention is low in cost andeasy to obtain; the preparation method disclosed by the invention is suitable for large-scale industrial production; and the prepared sulfate polymyxin B crystal product has the advantages of higher purity and clarity and low moisture absorption.

The invention belongs to the technical field of medicines, and in particular relates to a polymyxin E composition and a preparation method and application thereof. In the composition, total content of polymyxins E1, E2, E3, E1-1 and E1-7MOA is more than or equal to 80 percent, wherein the polymyxin E is preferably polymyxin E sulfate or polymyxin E sodium mesylate, and is particularly preferably polymyxin E sodium mesylate. The polymyxin E composition can be used for preparing medicines for treating Gram-negative bacterial infections.

The invention discloses a polymyxin sulfate B monoclonal antibody hybridoma cell strain and application thereof, and belongs to the field of food safety immunodetection. The polymyxin sulfate B monoclonal antibody hybridoma cell strain OVA17 is obtained through screening and is preserved in the China General Microbiological Culture Collection Center (CGMCC), and the preservation number is CGMCC No.18516. The monoclonal antibody secreted by the cell strain has good specificity and detection sensitivity (IC50 value is 13.13 ng / mL) for polymyxin B sulfate, can realize detection of polymyxin B sulfate residue in feed and livestock and poultry food, provides a raw material for immunodetection of polymyxin B sulfate residue in food. Practical application value is achieved.

The invention relates to a polymyxin derivative wherein the derivative has a total of three positive charges at physiological pH and wherein the terminal moiety (D) of the derivative comprises a total of 1 to 5 carbon atoms; and to a combination product comprising at least two such derivatives. The invention further relates to a method for sensitizing Gram-negative bacteria to an antibacterial agent by administering, simultaneously or sequentially in any order a therapeutically effective amount of said antibacterial agent and a derivative according to the invention to said subject; to methods for developing novel antibiotics; and for sensitizing clinically important bacteria to a host defense mechanism complement present in serum. The invention also relates to a method of treating a subject for a gram-negative bacterial infection by administering a polymyxin derivative of the invention in combination with a second antibacterial agent. Finally, the invention relates to a process for preparing such polymyxin derivatives.

Described are compounds of formula (I) for use in combination treatment with a second active agent, such as rifampicin, for example for treatment of a microbial infection. The compound of formula (I) is a polymyxin compound is:where the groups -A-, —R1, —R2, —R3, —R4, —R5, —R6, —R7, —R8, and —X— are described in detail within the description.

The invention discloses a polymyxin E component, a photochemical product thereof and a liquid chromatography-mass spectrometry analysis method, wherein the polymyxin E component comprises polymyxin Edouble-bonded components, and specifically comprises a polymyxin E1 double-bonded component, and a polymyxin E2 double-bonded component, and double bonds of these components are tested through high-performance liquid chromatography-mass spectrometry (HPLC-MS) and secondary mass spectrometry analysis to be located at the end of N-fatty acyl chain. The double-bonded components are subjected to photochemical reaction, namely a Paterno-Buchi reaction (reaction), under the condition of acetone serving as a reaction reagent to obtain the photochemical product, and the product is verified via HPLC-MSto be a cycloaddition product; the above double-bonded components are designated as 7'-alkenylpolymyxin E1 and 6'-alkenylpolymyxin E2. The polymyxin E double-bonded components may have antibacterialactivity or toxicity different from that of common polymyxin E components and has potential application value, and the structural confirmation for the compounds also provide a research basis for strictly controlling the quality of polymyxin E.

Monoclonal antibodies against bacterial proteins that confer resistance to the antibiotic colistin and hybridomas that produce such monoclonal antibodies are described. The monoclonal antibodies have a strong affinity for MCR proteins and may be used in detection methods and kits to detect the presence of MCR proteins including variants thereof in a sample.

The invention discloses a preparation method of colistin sulphate. In the preparation method, the colistin sulphate is prepared through fermentation liquor pretreatment, reduced-pressured concentration, heating, precipitation separation and spray drying by extracting colistin from fermentation liquor, and the total yield of the colistin sulphate reaches 75-85 percent. The preparation method disclosed by the invention has the advantages of high specifity and yield, short process, small equipment investment and low cost; and the environmental pollution is reduced because solvents are not adopted in the whole process.

The invention discloses a technology for extracting polymyxin B through a precipitation method. In the technology, by means of the precipitation method, a polymyxin B fermentation liquid is subjected to precipitation with oxalic acid, wherein a precipitated and separated solid is dissolved with addition of an alkaline substance and then is adsorbed by a weak acidic cation exchange resin. The weak acidic cation exchange resin is washed by purified water and is per-washed and desorbed with dilute sulfuric acid. A desorption solution is subjected to precipitation with an alkaline substance and acidic substances are dissolved. Solid-liquid separation is carried out with a liquid being subjected to decoloration and nano-filtration and then being dried to obtain the polymyxin B. The technology is short in required processes and is free of organic solvents. The polymyxin B is high in content and is suitable for industrialized production and application.

The present invention provides, in part, a Paenibacillus sp. isolate, designated Paenibacillus polymyxa JB05-01-1, as well as an anti-microbial agent obtained from the bacterium or cell culture supernatant thereof. Compositions, methods and uses are also provided.

The invention belongs to the technical field of medicine synthesis, and discloses a method of pure-solid-phase synthesis of polypeptide antibiotic Colistin. By use of the method, the synthetic process and procedure of the Colistin are simplified, the synthesis time is shortened, and the use amount of solvents is reduced.