23 results about "Human insulin" patented technology

The invention discloses a serum substitute compatible umbilical cord mesenchymal stem cell culture medium additive composition and application thereof. The serum substitute compatible umbilical cord mesenchymal stem cell culture medium additive composition is prepared from the following components in parts by mass: 15 to 25 parts of platelet-derived growth factor-BB; 15 to 25 parts of basic fibroblast growth factors; 8 to 12 parts of transforming growth factor-beta; and 8-12 parts of recombinant human insulin. The serum substitute compatible umbilical cord mesenchymal stem cell culture medium additive composition and the application thereof disclosed by the invention have the following beneficial effects: (1) the proliferation efficiency is remarkably improved; (2) the cell morphology and dryness can be well maintained; and (3) the compliance and the compatibility are good.

The invention provides an induced pluripotent stem cell culture medium and a culture method of induced pluripotent stem cells. The induced pluripotent stem cell culture medium comprises a basic culture medium, a composite additive A and a composite additive B, the composite additive A comprises 5-30 [mu] g / mL of recombinant human insulin growth factors, 1-5 ng / mL of recombinant human transforming growth factors, 50-200 ng / mL of recombinant human basic fibroblast growth factors, 7-20 [mu] g / mL of sodium selenite, 0.5-2 mg / mL of albumin, 50-70 [mu] g / mL of ascorbic acid and 5-20 [mu] g / mL of transferrin; the composite additive B is prepared from 50 to 300mg / mL of a Stock B solution, 0.5 to 5 percent (v / v) of a non-essential amino acid solution, 50 to 200ng / mL of DL-piperidinecarboxylic acid, 50 to 200mu g / mL of gamma-aminobutyric acid, 0.1 to 1 percent (v / v) of a lipid concentrated solution, 0.5 to 5 percent (v / v) of an L-alanyl-L-glutamine solution and 0.1 to 0.1 percent of mercaptoethanol. The culture medium for the induced pluripotent stem cells provided by the invention can ensure the self-renewal capability of the iPSCs in a long-term passage process; and the genetic stability of the iPSCs cells is optimized according to the genome integrity and pluripotent state.

Polypeptide antagonists of the human insulin receptor are provided. In some embodiments, a pharmaceutical formulation is provided, comprising an INSR antagonist as identified herein, and a pharmaceutically acceptable excipient. The formulation may be provided in a unit dose, e.g. a therapeutically effective dose. In methods of the disclosure, an effective dose of a an INSR antagonist as identified herein is administered to an individual having, or at risk of having, hyperinsulinemia, in a dose effective to stabilize, reduce or prevent clinical symptoms of the disease. The individual may be monitored for clinical indicia of disease before, during, and / or after administration.

The application discloses a preparation method of a human insulin purity standard substance, which comprises the following steps of raw material selection, qualitative identification, standard substance uniformity, stability test, value determination and uncertainty evaluation. The application uses commercially available high-purity human insulin as a raw material, and after purity analysis, qualitative identification, uniformity test and stability test, isotope-labeled Val, Leu and Phe are used as internal standards, and liquid chromatography-isotope dilution mass spectrometry is used for value determination, so that the interference and influence of possible isoleucine substitution leucine impurities, isomer impurities or other molecular weight protein impurities on the value determination result can be effectively excluded, and the value determination result is more accurate and reliable. The purity standard substance prepared by the application can be used for serum insulin standard substance quality value tracing, quality control of insulin measurement results in in-vitro diagnosis, related method confirmation and evaluation, and evaluation of repeatability, linearity and cross contamination rate of a closed light-emitting immune analyzer.

A pharmaceutical formulation presented, comprising an insulin or an insulin analog, and at least one protective excipient, in high concentration, selected from: a protective osmolyte, a surfactant, and a phenolic preservative. In embodiments, the insulin or insulin analog is one or more of: regular human insulin, insulin lispro, insulin aspart, insulin glulisine, insulin glargine, insulin detemir, or insulin degludec. In some embodiments, at least one of: the concentration of the protective osmolyte confers an osmolarity greater than 250 mOsm / L, the concentration of the surfactant is greater than 0.1% w / v, or the concentration of the phenolic preservative is greater than 0.4% w / v. In some embodiments, insulin API is dissolved and incubated for 20 to 40 minutes at high pH before the protective excipients are added.

This invention belongs to the field of stem cell culture medium technology, and provides a cell culture medium for enriching extracellular vesicles derived from stem cells and its preparation method. This invention disperses sodium hyaluronate or sodium hyaluronate-chondroitin sulfate sodium on a defined basic culture medium and grafts porous calcium alginate-gelatin microcarriers onto its surface, while limiting the content of recombinant human insulin, recombinant human transferrin, sodium selenite, and recombinant human serum albumin. This forms a culture system that combines a cell interaction interface with separable microcarriers, improving the enrichment stability of extracellular vesicles derived from stem cells, reducing the risk of solid-liquid separation residues and coupling agent residues, and solving the problem of balancing enrichment capacity with solid-liquid separation and grafting stability with biosafety. It is suitable for in vitro stem cell culture and extracellular vesicle preparation.

The application discloses a kind of serum-free epidermal melanocyte culture medium and its application, method includes by dissolving adenine, hepatocyte growth factor (HGF), glutamine, retinoic acid, vitamin E, nicotinic acid, sodium selenite, sodium hyaluronate and recombination human type XVII collagen are configured into initial culture medium;Preparation of melanocyte stimulating hormone, endothelin-3, transferrin, triiodothyronine, cholesterol, neuroregulin 1 (NRG1), 3-isobutyl-1-methyl purine (IBMX), dibutyryl cyclic adenosine monophosphate (db-cAMP), cholera toxin, hydrocortisone and recombination human insulin mother liquor;Step three: according to culture and concentration, a certain amount of mother liquor of step two is added to initial culture medium.The culture medium can effectively promote human epidermal melanocyte proliferation, melanin accumulation and help to promote human umbilical cord mesenchymal stem cells into human epidermal melanocyte, provide key advantage for obtaining functional melanocyte with high activity and therapeutic potential.

The invention discloses a method for improving killing activity of NK cells, and belongs to the technical field of cellular immunity. Carrying out segmented in-vitro culture on the separated NK cells by adopting a specific basic induction culture medium, an activation induction culture medium and an amplification culture medium, wherein recombinant human insulin and sodium selenite are added into the basic induction culture medium; ganoderma lucidum polysaccharides, eicosapentaenoic acid, IL-15 and IL-18 are added into the activation and induction culture medium. Salidroside, tea polyphenol, low molecular weight chitosan, IL-2 (interleukin-2), IL-21 (interleukin-21) and SCF (mesenchymal stem cells) are added into the amplification culture medium. According to the method, the killing ability of the NK cells obtained through in-vitro culture on tumor cells and the in-vitro proliferation effect of the NK cells are remarkably improved, the problems that in the prior art, the in-vitro proliferation efficiency of the NK cells is low, and cell activity is difficult to consider are solved, and a foundation is laid for clinical transformation of NK cell immunotherapy.

Novel derivatives of human insulin are described that have binding affinities for the IGF-1 receptor surprisingly comparable to human IGF-1, while having higher affinities for both isoforms of the insulin receptor than human insulin. The novel insulin derivatives have D-histidine or glycine at position B24, the B chain has glycine at the C-terminus at position B31 and tyrosine at position B32, and glutamic or aspartic acid at position B10. The derivatives of the invention can be used particularly for growth and proliferation stimulation in cell cultures or for treatment of neurological disorders as of Alzheimer's disease, Huntington's and Parkinson's diseases, a cognitive loss or symptoms of syndromic autism, since one derivative could mediate both insulin and IGF-1 function.

Provided are a differentiation-inducing culture medium for the transformation of adipose-derived mesenchymal stem cells into chondrocytes and the use thereof. A basal culture medium of the differentiation-inducing culture medium comprises a nutrient solution and an inducer. The nutrient solution comprises a blood substitute, a non-essential amino acid, β-mercaptoethanol, penicillin / streptomycin, dexamethasone, sodium pyruvate, ascorbic acid, sodium selenite, transferrin, recombinant human insulin, β-glycerophosphate and TGF-β1; and the inducer comprises cyanamide, delphinidin chloride and an extracellular matrix derived from adipose-derived mesenchymal stem cells. The components of the inducer, i.e. cyanamide and delphinidin chloride, can stimulate chondrocyte generation by means of up-regulating the expression of aggrecan and collagen II; and the component of the inducer, i.e. extracellular matrix (ADSC-ECM), can increase the adhesion between chondrocytes, enhance the signaling communication between chondrocytes, and promote the growth and metabolism of chondrocytes, thereby greatly improving the differentiation efficiency of adipose-derived mesenchymal stem cells into chondrocytes.

The application relates to an aqueous pharmaceutical formulation comprising 200-1000 U / mL [equimolar to 200-1000 IU human insulin] of insulin glargine.

The invention provides a DC-CIK cell culture medium. The DC-CIK cell culture medium comprises a DC culture medium, a DC maturation factor, a CIK culture medium, a CIK activating factor and a coating solution. The DC culture medium is prepared from IL-4, GM-CSF, recombinant human albumin, recombinant human transferrin, recombinant human insulin, cholesterol, copper sulfate, manganese chloride, ethanolamine, progesterone, palmitic acid, astragalus polysaccharide and DMEM / F12; the DC maturation factor comprises TNF-alpha, IL-1beta and a DMEM / F12 culture medium; the CIK culture medium is prepared from IMDM, lysine, polysaccharide nucleic acid, L-alanyl-L-glutamine, adenosine deaminase and glucose; the CIK activating factors comprise Anti-CD3, TNF (Tumor Necrosis Factor)-alpha, IL-1beta and DMEM (Dulbecco Modified Eagle Medium) / F12; the value-added rate and the cell viability of the DC-CIK cells cultured by the cell culture medium disclosed by the invention are remarkably improved.

The invention provides a monoclonal antibody for identifying proinsulin. A light chain CDR3 of the monoclonal antibody is shown as SEQ ID NO: 13, a light chain CDR1 of the monoclonal antibody is shown as SEQ ID NO: 14, a light chain CDR2 of the monoclonal antibody is shown as SEQ ID NO: 15, a heavy chain CDR3 of the monoclonal antibody is shown as SEQ ID NO: 16, a heavy chain CDR1 of the monoclonal antibody is shown as SEQ ID NO: 17, and a heavy chain CDR2 of the monoclonal antibody is shown as SEQ ID NO: 18. The monoclonal antibody is used for identifying human proinsulin. The monoclonal antibody can specifically recognize the specific epitope of the proinsulin, has the advantages of high recognition specificity, good affinity, high titer and the like, and has wide application value.

A method and vectors for controlling blood glucose levels in a mammal are disclosed. In one embodiment, the method comprises the steps of: treating the hepatocyte cells of a patient with a first, second or third vector, wherein the first vector comprises a promoter enhancer, glucose inducible regulatory elements, a liver-specific promoter, a gene encoding human insulin with modified peptidase and an albumin 3′UTR and lacks an HGH intron, wherein the second vector comprises an HGH intron, glucose inducible regulatory elements, a liver-specific promoter, a gene encoding human insulin with modified peptidase site and an albumin 3′UTR and lacks a promoter enhancer, wherein the third vector comprises an HGH intron, glucose inducible regulatory elements, a liver-specific promoter, a gene encoding human insulin with modified peptidase site, an albumin 3′UTR and a promoter enhancer and observing the patient's insulin levels, wherein the patient's insulin levels are controlled.

The present invention is generally directed to methods for measuring Insulin Resistance (IR) and for modulation of Type 2 Diabetes (T2D) onset treatment. In particular, diagnosis of IR is based on detection of different forms of disulfide bonds pairs that may form in insulin, affecting the normal insulin structure. Abnormal insulin is defined as any insulin that contains at least one of the forms of “abnormal” disulfide bond pairs. The methods herein measure human insulin isomers' concentration in a sample by utilizing HPLC or a monoclonal / polyclonal antibody to human insulin that contains “normal” or one or more “abnormal” disulfide bonds combinations. Some aspects of the invention also provide kits adapted for use in such methods. The human insulin isomers' concentration results are fed into a system to regulate the level of exogenous insulin needed to augment the level of normal insulin detected by the methods for a specific patient, or effectively regulate the amount of metformin, sulfonylureas, thiazolidinediones (PPARy agonists; glitazones), GLP-1s, SGLT-2 inhibitors and DPP-4 inhibitors or other IR treatment to be administered to the patient. The system and methods comprise a feedback loop to maintain a therapeutic level of insulin, metformin or other IR treatment and decrease the risk of hypoglycemia and other complications. The current invention has the advantage of being a self-referencing IR assessment by utilizing the measurements of “normal” insulin compared to “abnormal” insulin and not relying only on glucose measurements, Body Mass Index or other references included in current IR measurement methods. This leads to a much more responsive and faster treatment for IR compared to current approaches, which employ methods such as, Oral Glucose Tolerance Test (OGTT), which takes hours to perform.

The present invention discloses a method for crystallizing recombinant Human Insulin at lab and manufacturing scale in the presence of zinc chloride and sodium chloride mixture, higher concentration of organic solvent (IPA-19 to 25 million) and adjusting the pH to 5.0 at a faster rate (≤5 minutes). The method further comprises adopting procedures wherein the settling time is reduced and the holding temperature is altered in order to facilitate consistent protein crystal formation between 15 μm-30 μm and to increase the robustness of the process.

The application relates to the field of biotechnology, and discloses a culture medium system for maintaining and directionally inducing pluripotency of embryonic stem cells and application thereof. The culture medium comprises the following components in mass fractions: human insulin: 8-12 parts; tetramethylene diamine: 0.5-0.8 parts; luteinizing hormone: 0.2-0.5 parts; vitamin mixture: 1-2 parts; bovine serum albumin: 2-4 parts; sodium bicarbonate: 0.5-1.0 parts; antibiotic mixture: 0.2-0.4 parts; biotin: 1-2 parts; bone morphogenetic protein: 0.5-1.0 parts; and magnesium ethylenediaminetetraacetate: 0.1-0.2 parts. The application constructs a multi-factor regulation system, combines a signal inhibition, a three-dimensional support and a metabolic regulation module, realizes stable maintenance of pluripotency of embryonic stem cells, significantly improves culture uniformity, structural support and anti-differentiation capacity, and breaks through the technical bottleneck of a traditional system in terms of induction efficiency and stability.

The invention provides a culture medium and a method for carrying out suspension culture and amplification on pluripotent stem cells by utilizing a bioreactor. The culture medium for the pluripotent stem cells comprises a DMEM-F12 (Dulbecco's Modified Eagle Medium F12) culture medium, L-ascobic acid 2-phosphate trisomum salt, recombinant human insulin, Transferrin, Sodium Selenite, FGF2-G3 (Fibroblast Growth Factor 2), TGF (Transforming Growth Factor) beta 3, NRG1 (Nitrogen Regulated Growth Factor 1), HEPES (Hydroxyethyl Polyether Sulfonate), Lithium Chorides (LiCl), Polyethylene glycol (PEG) and Heparin Sodium salt. The culture medium and the culture method provided by the invention have the advantages of low cost, high cell amplification multiple and uniform 3D cell sphere morphology, can generate pluripotent stem cells on a large scale and maintain long-term subculture, provide technical guarantee for scientific research work and commercial production, and have wide application prospects.

The present invention relates to a nucleic acid molecule comprising a nucleotide sequence encoding a metabolic protein or part thereof, or a sequence having at least 90% sequence identity with said metabolic protein or part thereof, a human insulin-like growth factor II (IGFII) gene sequence, and a nucleotide sequence encoding at least one peptide that promotes cellular uptake or transendocytosis, the nucleotide sequence is inserted at a position between the coding amino acids 28 and 42 of mature IGFII of the IGFII gene sequence. The invention also relates to related virus particles, fusion proteins and uses thereof.

The invention is a pharmaceutical composition of human insulin or an insulin analog comprising citrate, treprostinil and stabilizing agents, which has a faster pharmacokinetic and / or pharmacodynamic effect than commercial formulations of existing insulin analog products and which is stable for commercial use.

The invention provides a preparation method of lincetinib (OSI-906; the present invention relates to novel salts and crystalline forms of [8-amino-1-(2-phenyl-7-quinolyl) imidazo [1, 5-a] pyrazine-3-yl]-1-methyl-cyclobutanol], and to the use thereof. These new forms of lincetinib provide increased solubility and an improved pharmacokinetic profile for pharmaceutical formulations for oral administration. Thus, the present invention enables an improved method for the treatment of human insulin-like growth factor-1 receptor (IGF-1R) and insulin receptor (IR) mediated conditions.

Provided are an ultra-long-acting insulin analog, a preparation method therefor, and a use thereof. The present invention comprises modification of an amino acid sequence of natural human insulin, and by means of dual fatty acid modification at a suitable site, prolongs the acting time of an insulin analog, thereby improving patient compliance. The present invention solves the technical problem in the prior art that the drug acting time of insulin analogs is unsatisfactory, uses a recombinant expression-based preparation method, and has a simple process route and low costs.

The present invention relates to a composition in the form of an injectable aqueous solution having a pH of 3.5 to 4.4 and comprising at least human insulin A21G and at least one prandial acting glucagon inhibitor. In one embodiment, the prandial acting glucagon inhibitor is selected from an amylin analogue, or an amylin receptor agonist, or a GLP-1 analogue, or a GLP-1 receptor agonist (GLP-1 RA). In one embodiment, the prandial acting glucagon inhibitor is an amylin analogue or an amylin receptor agonist. In one embodiment, the prandial acting glucagon inhibitor peptide is pramlintide. The present invention also relates to a process for obtaining human insulin A21G comprising at least the step of reacting human insulin A21G, B31R, B32R (insulin glargine) with rat carboxypeptidase B in a ratio of insulin / carboxypeptidase of 500 to 2000 at a pH of 7.5 to 8.5 and a temperature of 20 to 30 °C for 10 to 20 hours.