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Dosing regimens for neural stem cell proliferating agents and differentiating agents for the treatment of neurological disorders

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Dosing regimens for neural stem cell proliferating agents and differentiating agents for the treatment of neurological disorders


Effective dosing regimens for neural stem cell proliferating and differentiating agents, kits comprising effective dosing regimens for neural stem cell proliferating and differentiating agents, and uses thereof are provided herein. Such kits and methods can be utilized acutely or chronically to treat a neurodegenerative disease or condition. Furthermore, the compositions and methods can be used continuously or intermittently in various dosing regimens.
Related Terms: Neurodegenerative Disease

Browse recent Stem Cell Therapeutics Corp. patents - Calgary, CA
Inventors: Samuel Weiss, Christopher Gregg, Allen Davidoff, Joseph E. Tucker
USPTO Applicaton #: #20120270779 - Class: 514 77 (USPTO) - 10/25/12 - Class 514 


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The Patent Description & Claims data below is from USPTO Patent Application 20120270779, Dosing regimens for neural stem cell proliferating agents and differentiating agents for the treatment of neurological disorders.

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CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a divisional of U.S. application Ser. No. 11/687,302, filed Mar. 16, 2007, which claims priority to and the benefit of U.S. Provisional Application Ser. No. 60/783,500, filed on Mar. 17, 2006; U.S. Provisional Application Ser. No. 60/789,132, filed on Apr. 5, 2006; and U.S. Provisional Application Ser. No. 60/862,669, filed on Oct. 24, 2006, which are incorporated herein by reference in their entireties.

BACKGROUND

The development of techniques for the isolation and in vitro culture of multipotent neural stem cells (for example, see U.S. Pat. Nos. 5,750,376; 5,980,885; 5,851,832) significantly improved the outlook for the treatment of neurodegenerative diseases and conditions. It was discovered that fetal brains can be used to isolate and culture multipotent neural stem cells in vitro. Moreover, in contrast to the long held belief that adult brain cells are not capable of replicating or regenerating brain cells, it was found that neural stem cells may also be isolated from brains of adult mammals. These stem cells, either from fetal or adult brains, are capable of self-replicating. The progeny cells can proliferate or differentiate into any cell in the neural cell lineage, including neurons, astrocytes and oligodendrocytes. Therefore, these findings not only provide a source of neural cells which can be used in transplantations but also demonstrate the presence of multipotent neural stem cells in adult brain.

Certain agents, neural stem cell proliferating agents, have been found to increase the number of neural stem cells in vitro or in vivo. The mechanisms for such increase may include stimulating proliferation, inhibiting differentiation, and/or preventing death of the neural stem cells. Additional agents, stem cell differentiating agents, have been found to selectively enhance the production of neuronal precursor cells or glial precursor cells in vitro or in vivo. These proliferating and differentiating agents can thus be employed to increase and selectively enhance neurons and glial cells.

SUMMARY

Provided herein are effective dosing regimens for neural stem cell proliferating agents and differentiating agents, kits, and uses thereof. Such compositions of matter and methods can be utilized acutely (e.g., within days after neural injury or onset of neurologic symptoms) or can be used chronically (e.g., for a persisting neural injury or ongoing neurologic symptoms). Furthermore, the compositions and methods can be used continuously or intermittently.

Accordingly, a method for treating or ameliorating a neurodegenerative disease or condition in a mammal is provided. The method comprises administering to the mammal an effective amount of hCG or LH and an effective amount of EPO, wherein the hCG or LH is administered systemically in at least three doses, optionally by use of a kit. The hCG, LH, and/or EPO can be administered either continuously or intermittently. Further, the hCG or LH can be administered in a first treatment period and EPO can be delivered in a second treatment period. For example, hCG or LH can be administered intermittently on days 1, 3, and 5 of a first treatment period, then EPO can be administered continuously on days 1, 2, and 3 of a second treatment period.

Also provided herein is a further method for treating or ameliorating a neurodegenerative disease or condition in a mammal. The method comprises administering to the mammal an effective amount of hCG or LH in a first treatment period followed by an effective amount of EPO in a second treatment period, optionally by use of a kit. The hCG or LH can be delivered intermittently during the first treatment period and the EPO can be delivered continuously during the second treatment period. For example, hCG or LH can be administered intermittently on days 1, 3, and 5 of a first treatment period, then EPO can be administered continuously on days 1, 2, and 3 of a second treatment period.

In the methods and kits, the treatment periods may be, for example, at least three days. The treating methods can be repeated several times or many times with second, third, forth, fifth, etc. treating periods. The treating methods, whether administered once, twice, several, or many times, can take the form of one or more kits.

The details of methods and kits are set forth in the accompanying drawings and the description below. Other features, objects, and advantages of the methods and kits will be apparent from the description and drawings, and from the claims.

BRIEF DESCRIPTION OF FIGURES

FIG. 1 shows the effect on functional recovery of a rat subjected to a Middle Cerebral Artery occlusion (MCAo) stroke with increasing dosages of hCG with an intravenous (IV) administration of 1440 IU EPO per day following intramuscular (IM) administration of dosages of hCG.

FIG. 2 shows the differential effect on functional recovery of a rat subjected to a MCAo stroke, compared to untreated controls, of 440 IU of hCG with an IV administration of 1440 IU EPO per day, hCG alone, or EPO alone.

FIG. 3 is a graph indicating % tissue loss (compared to non-stroke hemisphere) in rats subjected to a MCAo stroke, compared to untreated controls, of 440 IU of hCG with an IV administration of 1440 IU EPO per day, hCG alone, or EPO alone.

FIG. 4 shows representative images of tissue loss in rats subjected to a MCAo stroke, compared to untreated controls, of 440 IU of hCG with an IV administration of 1440 IU EPO per day, hCG alone, or EPO alone.

FIG. 5 is a bar graph indicating serum hCG levels as measured after a third IM administration of hCG in Example 2.

FIG. 6 shows the result of six day subcutaneous prolactin infusions in male rats at 10, 15, and 20 times the concentrations used for intracerebroventricular infusions. The total number of bromodeoxyuridine positive (BrdU+) cells in the subventricular zone (SVZ) for 8 sections from each animal is presented. The greatest increase in SVZ proliferation levels was observed with the 15 times dose (170 μg/day for 6 days). (10 times=113 μg/day; 20 times=226 μg/day; Control=rat serum albumin only (RSA)). Significance relative to control: 10×=*p<0.05; 15×=**p<0.01; 20×=p<0.05; n=3 for all conditions; one way analysis of variance (ANOVA) with Tukey posthoc test.

FIG. 7 shows the results of prolactin dosing in male rats using single daily intraperitoneal injections. The total number of BrdU+ cells per section are presented for each dosing regime. (A) A small increase in SVZ proliferation was observed with high 3 day doses. (B) The most robust dosing condition for increasing SVZ proliferation levels used a low, 170 μg/day dose each day over 6 days. Significance is relative to RSA control. n=3; *p<0.05; **p<0.01; one-way ANOVA followed by a Tukey posthoc test.

FIG. 8 shows that single intramuscular injections of hCG on days 1, 3, and 5 post-stroke (stroke=day 0) trigger significantly increased proliferation in the forebrain SVZ. Significant increases in the number of Phospho-Histone H3 positive (pHH3+) cells per ventricle were observed in the 1000 μg dose condition (n=3; *p<0.05; one way ANOVA with Tukey posthoc). Images show the nuclear label Hoechst and pHH3 expression in the dorsolateral corner of the lateral ventricles in RSA pial strip control rats versus 1000 μg hCG dosed animals, note the increase in total cell number and pHH3 expression in SVZ of 1000 μg dosed animals.

FIG. 9 shows that single intramuscular injections of 1000 μg per day of hCG on days 1, 3, and 5 post-stroke (stroke=day 0) trigger increased neurogenesis in the forebrain SVZ. The number of doublecortin+neurons was quantified in the dosed animals and was doubled in the 1000 μg dose animals. (n=3; **p<0.01).

FIG. 10 shows the results of single intramuscular injections of hCG given daily for 7 days starting 24 hrs post-stroke (stroke=day 0). (A) The daily 330 μg/injection dosing regime significantly increased the number of proliferating (pHH3+ cells) in the SVZ relative to all other dosing conditions and controls (n=3; *p<0.01; one way ANOVA with Tukey posthoc). (B) Observation of the ischemic lesions in the motor cortex of dosed rats revealed that animals receiving the 330 μg/injection daily dosing regime demonstrated new tissue growth and filling in of the lesion site with a tissue plug.

FIG. 11 shows increased proliferation in the SVZ of 330 μg/injection daily hCG dosed animals as confirmed by counts of BrdU+ cells. The number of BrdU+ cells per ventricle was significantly increased in the 330 μg/injection condition relative to control and 100 μg/injection (p<0.01; n=3; one way ANOVA with Tukey posthoc analysis). These results further confirmed the increase in proliferation observed with pHH3 staining



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stats Patent Info
Application #
US 20120270779 A1
Publish Date
10/25/2012
Document #
13401323
File Date
02/21/2012
USPTO Class
514/77
Other USPTO Classes
514/97
International Class
/
Drawings
11


Neurodegenerative Disease


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