Polynucleotides encoding relaxin

A polynucleotide and relaxin technology, applied in the field of polynucleotides encoding relaxin, can solve problems such as efficacy problems

Active Publication Date: 2019-04-16
MODERNATX INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

A recombinant form of relaxin known as Serelaxin and marketed by Novartis has been shown to have low toxicity, however, its efficacy is questionable because it degrades so rapidly in the bloodstream

Method used

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  • Polynucleotides encoding relaxin
  • Polynucleotides encoding relaxin
  • Polynucleotides encoding relaxin

Examples

Experimental program
Comparison scheme
Effect test

example 1

[1681] Example 1: Manufacture of Polynucleotides

[1682] According to the present disclosure, the manufacture of polynucleotides and parts or regions thereof can be accomplished using the methods taught in International Application WO2014 / 152027 entitled "Manufacturing Methods for Production of RNA Transcripts", The contents thereof are incorporated herein by reference in their entirety.

[1683] Purification methods may include those taught in International Applications WO2014 / 152030 and WO2014 / 152031, each of which is incorporated herein by reference in its entirety.

[1684] Methods of detection and characterization of these polynucleotides can be performed as taught in WO2014 / 144039, which is hereby incorporated by reference in its entirety.

[1685] Characterization of polynucleotides of the present disclosure can be accomplished using a program selected from the group consisting of polynucleotide mapping, reverse transcriptase sequencing, charge distribution analysis...

example 2

[1686] Example 2: Chimeric polynucleotide synthesis

[1687] introduce

[1688] According to the present disclosure, two regions or portions of a chimeric polynucleotide can be joined or linked using triphosphate chemistry.

[1689] According to this method, a first region or portion of 100 nucleotides or less is chemically synthesized with a 5' monophosphate and a terminal 3' deOH or block OH. If the region is longer than 80 nucleotides, it can be synthesized as two strands for ligation.

[1690] If the first region or portion is synthesized as a non-positionally modified region or portion using in vitro transcription (IVT), then the 5' monophosphate can be subsequently converted and the 3' end capped.

[1691] Monophosphate protecting groups may be selected from any of those known in the art.

[1692] The second region or portion of the chimeric polynucleotide can be synthesized using chemical synthesis or IVT methods. IVT methods can include RNA polymerases that can...

example 3

[1705] Example 3: PCR for cDNA Production

[1706] 2x KAPA HIFI using Kapa BioSystems (Woburn, MA) TM HotStart ReadyMix for PCR procedures for cDNA preparation. This system includes 2x KAPA ReadyMix 12.5μl; forward primer (10μm) 0.75μl; reverse primer (10μm) 0.75μl; template cDNA-100ng; 2 0, diluted to 25.0 μl. Reaction conditions were 95°C for 5 minutes, and 25 cycles of 98°C for 20 seconds, then 58°C for 15 seconds, then 72°C for 45 seconds, then 72°C for 5 minutes, then 4°C to stop.

[1707] Using PURELINK from Invitrogen TM The PCR Micro Kit (Carlsbad, CA) was used to clean up reactions (up to 5 μg) according to the manufacturer's instructions. Larger reactions will require cleaning with a higher capacity product. After cleaning, use NANODROP TM Quantify the cDNA and analyze by agarose gel electrophoresis to confirm that the cDNA is the expected size. The cDNA is then submitted for sequencing analysis, followed by an in vitro transcription reaction.

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Abstract

The invention relates to mRNA therapy for the treatment of fibrosis and / or cardiovascular disease, mRNAs for use in the invention, when administered in vivo, encode human relaxin, isoforms thereof, functional fragments thereof, and fusion proteins comprising relaxin, mRNAs of the invention are preferably encapsulated in lipid nano particles (LNPs) to effect efficient delivery to cells and / or tissues in subjects, when administered thereto, mRNA therapies of the invention increase and / or restore deficient levels of relaxin expression and / or activity in subjects, mRNA therapies of the invention further decrease levels of toxic metabolites associated with deficient relaxin activity in subjects.

Description

[0001] Cross References to Related Applications [0002] This application claims the benefit under 35 U.S.C. § 119(e) of U.S. Provisional Application 62 / 338,470, filed May 18, 2016, which is hereby incorporated by reference in its entirety. Background technique [0003] Acute heart failure (AHF) is a sudden decline that occurs when the heart cannot pump enough blood to meet the body's cardiac demands. Signs and symptoms can include dyspnea, edema, and fatigue, which can lead to acute respiratory distress and death. AHF, as well as other cardiovascular diseases, can be caused by a deficiency of circulating relaxin. [0004] Relaxin is a 6000 Da heterodimeric polypeptide endocrine and autocrine / paracrine hormone that belongs to the insulin gene superfamily. Relaxin promotes angiogenesis and contributes to the repair of the vascular endothelium. It exerts its effects on musculoskeletal and other systems by binding its receptors in different tissues, a process mediated by diffe...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K9/51A61K31/7105A61K31/7088A61K38/22A61P9/00C07K19/00A61K47/42
CPCA61K38/00A61K9/0019A61K9/127C07K2319/00C07K2319/31A61K31/7105C07K14/64A61P9/00A61K38/1796A61K47/6811A61P9/04A61K9/5123A61K38/2221A61K48/0058A61K47/6929C07K16/26C12N15/52C12N15/62
Inventor 夏志南B.蒂乔N.布里安孔-埃里斯A.杜西斯S.德皮西奥托V.普雷辛亚克S.霍奇I.麦克费迪恩K.贝内纳托E.S.库马拉辛赫
Owner MODERNATX INC
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