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Methods for reducing viral load in hiv-1-infected patients

An HIV-1, virus technology, applied in chemical instruments and methods, botanical equipment and methods, applications, etc., can solve problems such as untested CCR5 coreceptor inhibitors

Inactive Publication Date: 2009-07-29
原基因药物有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, previous studies have not examined different classes of CCR5 coreceptor inhibitors, such as combinations of anti-CCR5 mAbs and non-antibody CCR5 antagonists

Method used

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  • Methods for reducing viral load in hiv-1-infected patients
  • Methods for reducing viral load in hiv-1-infected patients
  • Methods for reducing viral load in hiv-1-infected patients

Examples

Experimental program
Comparison scheme
Effect test

Embodiment approach

[0087]The present invention provides a method for reducing HIV-1 viral load in a human subject infected with HIV-1, which comprises administering an effective HIV-1 viral load reducing dose to said subject at predetermined time intervals ( a) Humanized antibody PRO 140, or (b) anti-CCR5 receptor monoclonal antibody, said humanized antibody PRO 140 or anti-CCR5 receptor monoclonal antibody (i) and CD4+ in the subject CCR5+ cells bind and inhibit the fusion of HIV-1 and the cells, (ii) inhibit the fusion of HIV-1 and CD4+CCR5+ cells, the efficacy is equal to or higher than that of PRO140, (iii) without reducing the body of the subject Under the condition of the number of CD4+CCR5+ cells, covering the subject’s CD4+CCR5+ cells, and / or (iv) without inducing an increase in the plasma concentration of the subject’s circulating beta chemokine, and the subject’s CD4+CCR5+ cell combination, where PRO140 includes (i) two light chains, each light chain contains the expression product of plas...

Embodiment 1

[0210] Example 1: Combination testing of PRO 140 and HIV-1 invasion inhibitor in a fluorescent RET assay

[0211] Materials and methods

[0212] Compound and mAb

[0213] PRO 140 was prepared by expressing in Sp2 / 0 cells using hybridoma serum-free medium (Invitrogen, Carlsbad, CA) supplemented with 2mM L-glutamine. A 5.0 μm Depth filter (Sartorius, Goettingen, Germany) was used to purify a large amount of mAb, and then passed through a 0.2 μm sterilization grade filter (Sartorius). The mAb was purified by passing through an affinity column (MabSelect Protein A column, Amersham, Piscataway, NJ) and then by ion exchange chromatography (SP Sepharose Cation Exchange Resin, Amersham). Use Viresolve TM 10 Opticap NFP capsules (Millipore, Billerica, MA) followed by 0.2 μm filter nanofiltration PRO 140, and concentrated / diafiltered PRO 140 through a disposable TFF box (Millipore). Then, the mAb was purified through a hydroxyapatite column (Bio-Rad, Hercules, CA), concentrated to 10 mg...

Embodiment 2

[0298] Example 2: In the determination of HIV-1 pseudoviral particles (HIV-1PP), PRO 140 and small molecules, peptides and Protein inhibitor, combined test with HIV-1

[0299] Materials and methods

[0300] Preparation of HIV-1 fake particles

[0301] Through HIV-1 based NL4 / 3luc+env-plasmid and encoding HIV-1 JRFL Transient co-expression of Env constructs produced HIV-1 pseudoparticles (HIV-1pp) in 293T cells. NL4 / 3luc+env-plasmid was obtained from NIH AIDS research and reference reagent program (Cat.No.3418), and HIV-1 JRFL Env was inserted into the pcDNA3.1 vector (Invitrogen). Briefly, 293T cells were transfected with NL4 / 3luc+env-receptor vector and Env expression vector (Profection Mammalian Transfection Kit, Promega) calcium phosphate in a ratio of 1:1 in Hepes buffer. After 16 hours of transfection, the medium was aspirated, and fresh cell culture medium (DMEM containing 10% FBS, glutamine and antibiotics) was added, and the culture was continued at 37°C for another ...

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Abstract

This method provides a method for reducing HIV-I viral load in an HIV-1-infected human subject which comprises administering to the subject at a predefined interval effective HIV-I viral load- reducing doses of (a) a humanized antibody designated PRO 140, or of (b) an anti-CCR5 receptor monoclonal antibody. This invention also provides a method for inhibiting in a human subject the onset or progression of an HIV-I -associated disorder, the inhibition of which is effected by inhibiting fusion of HIV-I to CCR5CD4 target cells in the subject. This invention also provides a method for treating a subject infected with HIV-I comprising administering to the subject (a) a monoclonal antibody which (i) binds to a CCR5 receptor on the surface of the subject's CD4 cells and (ii) inhibits fusion of HIV-I to the subject's CCR5CD4 cells, and (b) a non-antibody CCR5 receptor antagonist, in amounts effective to treat the subject.

Description

[0001] This application claims the rights of the following applications: U.S. Provisional Application No. 60 / 702,064 filed on July 22, 2005; U.S. Provisional Application No. 60 / 701,889 filed on July 23, 2005; U.S. Provisional Application No. filed on August 26, 2005 Provisional Application No. 60 / 711,528; and U.S. Provisional Application No. 60 / 715,619 filed on September 9, 2005; the entire content of each article is hereby incorporated into this application by reference. [0002] The present invention was completed with the support of U.S. government fund numbers AI046871 and AI066329, which came from the National Institute of Allergy and Infectious Diseases. Therefore, the US government has certain rights in the subject invention. [0003] Throughout this application, multiple publications are mentioned in parentheses by author name and date, or by patent or patent publication number. The full citations of these publications can be found at the end of the description immediately p...

Claims

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Application Information

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IPC IPC(8): A61K39/42A01N61/00C07K16/00
Inventor 威廉·C·奥尔森保罗·J·马多恩丹尼尔·C·佩韦尔罗伯特·J·伊斯雷尔乔斯·D·穆尔格阿
Owner 原基因药物有限公司
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