73 results about "Adenylyl Cyclases" patented technology

This invention relates to modified Vasoactive Intestinal Peptide (VIP) / Pituitary Adenylate Cyclase Activating Peptide (PACAP) Receptor 2 (VPAC2) agonists (VPAC2 agonists) comprising a VPAC2 agonist linked to a polyethylene glycol polymer, as well as related formulations, dosages, and methods of administration thereof for therapeutic purposes. These VPAC2 agonists, compositions, and methods are useful in providing a treatment option for those individuals afflicted with a metabolic disorder such as diabetes, impaired glucose tolerance, metabolic syndrome, or prediabetic states, by inducing glucose-dependent insulin secretion in the absence of the therapeutically limiting side effect of reducing or lowering blood pressure.

This invention provides novel peptides that function in vivo to stimulate insulin release from pancreatic beta cells in a glucose-dependent fashion. These insulin secretagogue peptides are shown to stimulate insulin release in rat islet cells in vitro, and in vivo. The peptides of the present invention provide a new therapy for patients with decreased endogenous insulin secretion, in particular type 2 diabetics. In particular, the invention is a polypeptide selected from a specific group of VIP / PACAP-related polypeptides, or functional equivalents thereof. The invention is also directed to a method of treating a metabolic disease in a mammal comprising administering a therapeutically effective amount of the insulin secretagogue peptides to said mammal. Also disclosed are methods of making the peptides, both recombinant and synthetic.

The present invention is directed to novel methods of treating a subject with a bone deficit disorder. The methods generally include administering to a subject in need thereof a pharmaceutically acceptable formulation comprising a parathyroid hormone (PTH) peptide analogue in a daily dose of 2 μg to 60 μg, wherein said PTH peptide analogue has a reduced phospholipase-C activity and maintains adenylate cyclase activity.

This invention relates to novel analogs of pituitary adenylate cyclase-activating polypeptide (PACAP), which are agonists for the PACAP / vasoactive intestinal peptide (VIP) receptors: PAC1, VPAC1 and VPAC2 receptors. These PACAP analogs can be used as prophylactic / therapeutic agents for a wide range of medical disorders, including (but not limited to) cancer and autoimmune disease. These PACAP analogs can be coupled to suitable radionuclides and used in the localization, diagnosis and treatment of disseminated cancers and metastatic tumors, or coupled to small molecule therapeutics and used as vectors for targeted drug delivery. This invention also provides pharmaceutical compositions of one or more PACAP-like compounds of the invention either alone or in combination with one or more other prophylactic / therapeutic agents.

The present invention is directed to novel methods of treating a subject with a bone deficit disorder. The methods generally include administering to a subject in need thereof a pharmaceutically acceptable formulation comprising a parathyroid hormone (PTH) peptide analogue in a daily dose sufficient to result in an effective pharmacokinetic profile and maintained adenylate cyclase activity, while simultaneously reducing undesirable side effects.

The invention provides adenylate cyclase, and a coding gene, a vector, a bacterial strain and application thereof. A DNA (Deoxyribose Nucleic Acid) sequence for coding the adenylate cyclase provided by the invention has a base sequence represented by SEQ ID NO.:1. Furthermore, the invention further provides the adenylate cyclase coded by the DAN sequence, a recombinant vector including the DAN sequence, a host cell including the recombinant vector, and applications of all in production of the adenylate cyclase. The genetic engineering strain of colibacillus, disclosed by the invention, can beused for expressing the adenylate cyclase with high efficiency. The inducible enzyme activation thereof can achieve 7 U / mg or 10 U / mg. The bacterial strain is used for producing cyclic adenosine monophosphate and has the advantages of simple process, moderate condition, short cycle, few by-products and the like.

The present invention relates to a naturally occurring low molecular weight adenosine A3 receptor agonist (LMW-A3RAg) which is preferably obtained from a vertebrate tissue or a vertebrate-derived cell by extraction in a liquid medium. The LMW-A3RAg of the invention is characterized by the following feature: (i) it is obtainable from animal-derived tissue or cells; (ii) it filters through a filter with a maximal molecular weight cut-off of about 3,000 Daltons; (iii) it is water soluble, heat stable, non-proteinaceous and resistant to adenosine deaminase activity. The invention also concerns pharmaceutical compositions comprising the naturally occurring LMW-A3RAg of the invention and therapeutic methods comprising administering to a subject in need an effective amount of the naturally occurring A3RAg for achieving a therapeutic effect, the therapeutic effect comprises inhibition of adenylate cyclase in target cells.

The present invention relates to a method of preventing or treating a disease caused by bacterial infection by administering an effective amount of a modulator of bacterial adenylyl cyclase. The invention also provides pharmaceutical compositions useful for preventing or treating a disease, with the compositions containing a therapeutically effective amount of a modulator of bacterial adenylyl cyclase. The invention also provides screening methods for identifying selective modulators of bacterial adenylyl cyclase that do not substantially modulate adenylyl cyclase of the subject. The invention also provides methods for culturing bacterial pathogens and methods for inducing the pathogenic state in vitro.

Soluble adenylyl cyclase (sAC) is implicated in proliferation of keratinocytes. Inhibitors of sAC are useful for the treatment and / or prevention of psoriasis and other hyperproliferative skin disorders. Assays to identify such compounds are also described.

The invention disclosed herein provides for methods of treating cancer using inhibitors of the Raf / Mek / P-Erk 1 / 2 pathway. These inhibitors include B2AR agonists (such as ARA-211 (pirbuterol) and isoproterenol), adenylyl cyclase activators, cAMP analogs and Epac activators. The invention also provides methods for diagnosing cancer in an individual.

The invention aims to proliferate or establish undifferentiated pluripotent stem cells that retain their differentiation potency by culturing pluripotent stem cells in a medium free of a feeder cell, or a serum. The aim is attained by using a culture medium for pluripotent stem cells comprising the known ingredients, which is supplemented with an inhibitor of an adenylate cyclase activity.

The invention provides a method for enhancing the ectogenesis of sheep oocyte. The method adopts an adenylate cyclase activator Forskolin (FSK), phosphodiesterase inhibitor cilostamide (CIL) and 3-isobutyl-1-methylxanthine (IBMX), treats the sheep oocyte in a combining manner and improves the capability of the ectogenesis of the sheep oocyte. The method comprises the following detailed steps of: picking the ovary of the killed sheep, putting the ovary of the killed sheep into normal saline for cleaning for 3-4 times; pumping the 2-8mm follicles on the surface of the ovary by using a 10ml syringe which absorbs 1ml ovum-pumping liquid containing 100mu mol / L FSK and 500mumol / L IBMX and is provided with a number-9 needle head, and injecting the follicular fluid recovered in the syringe into a35-60mm vessel; picking the oocyte under a microscope, putting the oocyte into the pumped follicular fluid, cleaning the oocyte for three times with in-vitro mature fluid containing 20mu mol / L CIL, and putting the oocyte in a CO2 incubator for culture; then cleaning the oocyte for three times with in-vitro mature fluid which does not contain CIL, putting the oocyte in the CO2 incubator for culture and adding into in-vitro fertilization fluid drop; unfreezing the seminal fluid with water bathing, and commonly incubating with the oocyte; and calculating the cleavage rate after 24 hours and calculating the blastocyst rate after 168 hours.

The present invention includes the discovery of a strain of Mycobacterium comprising an expression vector encoding a di-adenylate cyclase enzyme. The Mycobacterium is selected from the group consisting of Mycobacterium tuberculosis, Mycobacterium bovis, or a combination thereof and the preferred strain of Mycobacterium is BCG. The preferred expression vector is a mycobacterial expression vector including an hsp60 promoter and a DNA sequence of diadenylate cyclase (disA), or a functional part thereof. The strains of Mycobacterium are used in therapeutic applications including tuberculosis and cancer.

The invention discloses a method for preparing cyclic adenosine monophosphate by adenylate cyclase. The method comprises the following steps that the adenylate cyclase serves as a catalyst, adenosinetriphosphate serves as a substrate, and in the presence of Mg <2+>, a reaction solution containing the cyclic adenosine monophosphate is obtained. According to the adenylate cyclase, an amino acid sequence as shown in SEQ ID NO.14 is provided, the stability is good, the half-life period is long at medium and low temperatures, ATP can be efficiently catalyzed to generate cAMP in one step at mediumtemperature, a substrate conversion rate reaches 90 % or above, the advantages of simple industry, mild conditions, short period, few by-products and the like are achieved, the clean and pollution-free effects are achieved, and the energy conservation, consumption reduction and emission reduction in the cAMP production process can be realized.

The invention provides cells and methods for identifying modulators of signal transduction, based on transducisome proteins that coordinate and assemble many types of signal transduction proteins. A transducisome is a PDZ domain containing protein that binds at least one signal transduction protein or a PDZ domain containing protein with at least one signal transduction protein bound. Examples of transducisome proteins include INAD, GRIP and other recently identified multi-PDZ domain proteins. Examples of signal transduction proteins include GPCRs, tyrosine kinase receptors, tyrosine phosphatase receptors, ion channels, phospholipases, adenylate cyclases, kinases and G-proteins. Also provided are methods for identifying modulators of signal transduction, proteins (and polynucleotides encoding the same) corresponding to transducisomes, modified transducisomes or defective transducisomes to use in assays of signal transduction, and a screening assay system for detecting protein-protein interactions.

The invention provides methods for treating, ameliorating or protecting (preventing) an individual or a patient having or at risk of having heart disease or heart failure, or decreased cardiac function, comprising: providing a cyclic adenosine monophosphate-incompetent (cAMP-incompetent) adenylyl cyclase type 6 (AC6) protein or polypeptide (also called "an AC6mut"), or an AC6mut -encoding nucleic acid or a gene operatively linked to a transcriptional regulatory sequence.

An immunogenic composition comprising a recombinant protein comprising a Bordetella CyaA, or a fragment thereof, and a peptide that corresponds to a tumor antigen is provided as a cancer treatment. Methods of treatment with this immunogenic composition are also provided. In an embodiment, the therapeutic composition is a treatment for melanoma, and comprises epitopes from the HLA*0201 epitope. These epitopes include Tyr or GnT-V, and are present in the recombinant proteins CyaA-E5-Tyr and CyaA-E5-GnT-V.

The invention discloses a qualitative and quantitative determination method for active proteins in a pertussis toxin product and a diphtheria-pertussis-tetanus vaccine. A high-performance liquid chromatography-tandem mass spectrometry method is adopted for establishing the high-flux high-selectivity high-sensitivity quantitative method for active proteins including pertussis toxin, filamentous hemagglutinin, adhesin, mycoprotein and adenylate cyclase toxin in the pertussis toxin product and the diphtheria-pertussis-tetanus vaccine. According to the method, characteristic peptide fragments which can be used for qualitative and quantitative analysis of various active vaccine proteins are screened out from a complex vaccine matrix for the first time, and the peptide fragments are different from other reported protein peptide fragments, cannot be obtained through a protein search library and cannot also be simply obtained through amino acid sequences of the vaccine proteins. Simultaneous quantification of five pertussis toxin subunits, filamentous hemagglutinin, adhesin, mycoprotein and adenylate cyclase toxin in pertussis toxin products and diphtheria-pertussis-tetanus vaccines from different manufacturers and of different batches can be realized.

Provided are compositions for long-term maintenance of functional hepatocytes in culture, a method for improved maintenance of functional hepatocytes in vitro, and functional hepatocytes cultures according to the methods. The culture compositions include at least: one activator of adenylate cyclase, one TGFbeta inhibitor, one Notch inhibitor, one Wnt inhibitor, and / or one BMP inhibitor. The combinations of compounds are added to any hepatocyte cell culture medium in an effective amount to maintain functional hepatocyte function in vitro, long term. The hepatocytes can be used for in vitro drug research and to model liver disease.

The invention discloses a natural medicine composition for treating involutional depression and non-classical depression and use of the natural medicine composition, and belongs to the technical field of research of natural medicine formulas in compatibility dosage effect and use of the natural medicine. The composition consists of the components in percentage by mass: 70%-97.5% of hippophae rhamnoides fruit oil and 2.5%-30% of pseudo-ginseng stem and leaf total saponins. The natural medicine composition disclosed by the invention is reasonable in compatibility and safe in dosage, is free from any side effects on human bodies, can solve the problems that a conventional medicine has side effects and poor treatment effects when being used for treating climacterium and involutional depression, and has a significant regulation effect on serum marker indexes, namely adenylate cyclase-cyclic adenosine monophosphate and cyclic guanosine monophosphate, and reproductive hormonesm and target acceptors of various impact factors, such as follicular generation promoting hormones, pitocin, dihydrotestosterone hormones, female hormones, adrenocorticotropin hormones, cytokine interleukin-6, interleukin-8 cytokine and interleukin-2, so as to achieve a treatment effect on treatment of the involutional depression and the non-classical depression diseases.

The invention discloses a preparation method of purified coupling immobilized adenylate cyclase. The preparation method comprises the following steps: (1) preparing an affinity adsorption medium: adding 5-100g of a metal chelating resin carrier with surface modified by iminodiacetic acid to 10-200mL of 0.01-1mol / L of an NiCl2.6H2O solution, conducting oscillating at 25-30 DEG C and under 150-200rpm for 2-4h, and conducting filtration, so that the affinity adsorption medium is obtained; (2) immobilizing enzyme: adding 1.5-15ml of an adenylate cyclase solution which is 0.1-2mg / mL in concentration to 0.1-1g of the affinity adsorption medium obtained in the step (1), conducting oscillating at 25-30 DEG C and under 150-200rpm for 2-4h, and conducting centrifuging, so that the immobilized adenylate cyclase is obtained, wherein the adenylate cyclase is provided with a histidine tag. The immobilized enzyme prepared by the invention has the advantages of being high in enzymatic activity, low in enzymatic activity loss and the like, and the immobilized enzyme can be repeatedly used.

The present invention relates to methods and compositions for the treatment, management, or prevention of multiple myeloma and / or renal dysfunction in mammals. The methods of the invention comprise the administration of an effective amount of one or more pituitary adenylate cyclase activating polypeptide (“PACAP”) compounds, which includes PACAP, vasoactive intestinal peptide (“VIP”), their agonists, analogs, fragments, or derivatives, having one or more PACAP activities. The invention also provides pharmaceutical compositions comprising one or more PACAP compounds of the invention either alone or in combination with one or more other prophylactic / therapeutic agents useful in therapy for the treatment, management, or prevention of multiple myeloma and / or renal dysfunction.

The invention discloses a method for improving adenylate cyclase expression activity and increasing cAMP content in plants. The method comprises the following steps: selecting a template DNA sequencecapable of encoding a catalytic activity center of adenylate cyclase by using biological information resources, adopting PCR amplification or chemical synthesis or other selectable cloning methods toobtain a DNA product with the sequence fragment, and after transcriptional expression of the DNA product, generating a peptide chain or protein with adenylate cyclase catalytic activity by translation; introducing the obtained DNA product into a selected plant gene expression vector system by using a DNA cloning technology, and inserting the plant gene expression vector system into multiple cloning sites between promoter and terminator element sequences for controlling exogenous gene expression; and carrying out genetic transformation on the constructed plant gene expression vector to obtain atransgenic plant with significantly improved adenylate cyclase expression activity and significantly increased cAMP content. The method provided by the invention provides a new idea for people to develop and utilize cAMP in plants, and fills the blank of the research.

The invention relates to the technical field of cell engineering, in particular to a culture medium for directional induced differentiation of pluripotent stem cells into hepatocytes, a culture method and application. The invention provides a culture medium composition, the culture medium composition contains an adenylate cyclase activator, an ALK5 inhibitor, a growth factor, OSM and other factors, and the maturity of hepatocytes obtained by induced differentiation of iPSC can be greatly improved by using the culture medium. The ASGR1 + cell proportion is increased, and the ALB secretion level is increased. A reporter gene is added into an ASGR1 gene by utilizing a gene editing and recombination technology, a vector containing the two genes is introduced into the pluripotent stem cell, and after the pluripotent stem cell is induced to be differentiated into the hepatocyte, the reporter gene and the ASGR1 are co-expressed, so that the proportion of ASGR1 + hepatocyte subpopulation in differentiated cells is favorably detected, the ASGR1 + cell population is detected or sorted by utilizing FACS, and the detection accuracy of the pluripotent stem cell is improved. Extra experimental operation is not needed, the cells can be directly detected or sorted on a machine after being digested, and high-throughput detection can be realized by utilizing the detection method disclosed by the invention.

The invention discloses adenylate cyclase containing an FYVE structural domain from phytophthora sojae as well as an encoding gene and application of the adenylate cyclase. The adenylate cyclase is prepared from phytophthora sojae. The adenylate cyclase PsZFAC containing the FYVE structural domain has an amino acid sequence as shown in SEQ ID No. 4, and a coding gene of the adenylate cyclase PsZFAC has a nucleotide sequence as shown in SEQ ID No. 3. The main function of the PsZFAC protein is to control the formation of phytophthora sojae sporangium. The gene provided by the invention has a very high application value in controlling phytophthora sojae root rot, and a bactericide developed by taking the protein as an action target has important practical significance in controlling the occurrence and prevalence of phytophthora sojae root rot.