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Enolase 1 (ENO1) compositions and uses thereof

a technology of enolase and composition, applied in the field of enolase 1 (eno1) composition, can solve the problems of reducing the quality of life or even being fatal, deregulation of glucose metabolism, and short serum half-life of insulin, and achieves the effect of improving blood glucose level control, and increasing glucose flux in the skeletal muscle cell of the subj

Inactive Publication Date: 2019-10-10
BERG LLC +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

Eno1 treatment effectively increases glucose uptake and insulin sensitivity, normalizes glucose tolerance, and treats diabetes by enhancing glucose flux and glycolytic activity in skeletal muscle cells, thereby addressing the limitations of existing diabetes management strategies.

Problems solved by technology

Because these conditions can reduce the quality of life or even be fatal, strategies to restore adequate glucose clearance from the bloodstream are required.
The consequent reduction in insulin production inevitably leads to the deregulation of glucose metabolism.
While the administration of insulin provides significant benefits to patients suffering from this condition, the short serum half-life of insulin is a major impediment to the maintenance of normoglycemia.
An alternative treatment is islet transplantation, but this strategy has been associated with limited success.
Although various therapeutic treatments are available for the management of type 2 diabetes, they are associated with various debilitating side effects.
Such lifestyle changes, however, are not sufficient to reverse the vascular and organ damages caused by diabetes.

Method used

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  • Enolase 1 (ENO1) compositions and uses thereof
  • Enolase 1 (ENO1) compositions and uses thereof
  • Enolase 1 (ENO1) compositions and uses thereof

Examples

Experimental program
Comparison scheme
Effect test

example 1

Platform Technology to Identify Enolase 1 (Eno1) as an Important Node of Activity in the Etiology of Diabetes

[0482]In this example, the platform technology described in detail in international Patent Application No. PCT / US2012 / 027615 was employed to integrate data obtained from a custom built diabetes model, and to identity novel proteins / pathways driving the pathogenesis of diabetes, particularly type 2 diabetes. Relational maps resulting from this analysis have identified Eno1 as an important node of activity in the etiology of diabetes. Therefore, Eno1 is an important diabetes treatment target, as well as a diagnostic / prognostic marker associated with diabetes.

example 2

lation of Glucose Uptake in Myotubes

[0483]Eno1 was recombinantly expressed in E. coli as a 6×HIS protein tag using a commercially available expression vector. The tagged Eno1 was purified using affinity chromatography methods known in the art. Preferably, the 6×HIS tag was cleaved to produce the protein for use in the methods provided herein.

[0484]Human skeletal muscle myoblasts (HSMM) were procured from PromoCell and were cultured in growth media recommended by the vendor. HSMM myoblasts (20,000 cells / well) were differentiated with 2% horse serum in 96 well plates for 7 days before experiment. Cells were treated with human Eno1 (500 ug / ml). Cells were washed twice with 200 μl MBSS modified balanced salt solution (MBSS) buffer containing 0.1% BSA, and then serum starved with 100 ul MBSS 0.1% BSA for 4 hours. Upon initiation of insulin stimulation, 100 ul 2× reagents in MBSS 0.1% BSA buffer was added to 100 ul starvation media to make 1× concentration for the experiment. The 2× reage...

example 3

els of Diet Induced Obesity (DIO) Mice

[0489]Two essentially equivalent models of diet induced obesity were used in the methods provided herein.

[0490]In the first method, male C57BUL6J mice were obtained from Jackson Laboratories (Bar Harbor, Me.) and initially housed 4-5 per cage at 22° C. on a 12:12 hr day-night cycle. Beginning at 6 weeks of age, mice were fed with a high-fat diet (Research Diets Cat #: D12492; 60 kcal % fat, 20 kcal % protein, and 20 kcal % carbohydrate). Lean control mice were also obtained and fed a standard diet. Body weight of DIO mice before experiments was significantly heavier than that of lean control mice. In one study, DIO mice weighed 38.4±0.6 g whereas lean mice weighed 29.9±0.5 g (p<0.05).

[0491]In the second method, diet induced obese male C57BU6J mice (12 week old) and control lean mice (12 week old) were obtained from Jackson Laboratories (Bar Harbor, Me.) and initially housed 4-5 per cage at 22° C. on a 12:12 hr day-night cycle. Mice were acclimat...

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PUM

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Abstract

The invention provides compositions comprising Eno1 for delivery to a muscle. Further, the invention provides a method for normalizing blood glucose in a subject with elevated blood glucose, comprising administering to the subject enolase 1 (Eno1), thereby normalizing blood glucose in the subject. The invention also provides methods of treating one or more conditions including impaired glucose tolerance, insulin resistance, pre-diabetes, and diabetes, especially type 2 diabetes in a subject, comprising administering to the subject enolase 1 (Eno1), thereby treating the condition in the subject. In certain methods of the invention, the Eno1 is delivered to muscle.

Description

RELATED APPLICATIONS[0001]This application is a continuation of U.S. patent application Ser. No. 14 / 596,207, filed Jan. 13, 2015 which, in turn, claims priority to U.S. Provisional Patent Application No. 61 / 926,913 filed on Jan. 13, 2014, U.S. Provisional Patent Application No. 62 / 009,783 filed on Jun. 9, 2014, and U.S. Provisional Patent Application No. 62 / 100,881 filed on Jan. 7, 2015, the contents of each of which are incorporated herein in their entirety.SUBMISSION OF SEQUENCE LISTING[0002]The Sequence Listing associated with this application is filed in electronic format via EFS-Web and hereby incorporated by reference into the specification in its entirety. The name of the text file containing the Sequence Listing is 119992_10607_Sequence_Listing. The size of the text file is 58 KB, and the text file was created on Nov. 27, 2018.BACKGROUND[0003]As the levels of blood glucose rise postprandially, insulin is secreted and stimulates cells of the peripheral tissues (skeletal muscl...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): A61K38/51A61K47/64A61K47/59A61K9/00A61K49/00C12N9/88C12Q1/54A61K38/00
CPCC12N9/88G01N2800/042A61K47/64A61K47/59C12Y402/01011G01N2333/988A61K38/51A61K49/0008C12Q1/54A61K38/00A61K9/0019A61K9/0053A61P3/08A61P43/00A61P5/50A61P3/10
Inventor NARAIN, NIVEN RAJINSARANGARAJAN, RANGAPRASADVISHNUDAS, VIVEK K.GESTA, STEPHANEJING, ENXUANDAUNERT, SYLVIADEO, SAPNA K.JIMENEZ, JOAQUIN J.DIKICI, EMREDAFTARIAN, PIROUZ MOHAMMAD
Owner BERG LLC
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