IP Library › Granted Patent US 12,421,494
Granted Patent B2
US 12,421,494 · App. 18/584,070 · Granted Sep 23, 2025

Method for ex-vivo expansion of regulatory T cells with enhanced suppressive function for clinical application in immune mediated diseases

Inventors: Tinghua Cao (Malvern, PA); Li Li (Downington, PA)
Assignee: Therakos Development Limited
C12N5/0637A61K40/11A61K40/22A61K40/416C12N5/0636A61K2035/122C12N2501/23C12N2501/2302C12N2501/51C12N2501/515
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Quick Facts
Patent No.
US 12,421,494
App. No.
18/584,070
Granted
Sep 23, 2025
Kind
B2
Abstract

The invention provides methods for the ex-vivo expansion of CD4+CD25+ Tregs. The invention provides a method for producing ex vivo expanded Tregs that may be used to inhibit unwanted human immune responses against self-antigens or allergens. Additionally, the ex vivo expanded Tregs may provide treatment for inflammatory/autoimmune diseases.

Claims (16)

1. A method of preparing a population of cells, the method comprising:

a) obtaining a population of T cells from an individual with an immune-mediated disease selected from Systemic Lupus Erythematous, Multiple Sclerosis, asthma, Rheumatoid Arthritis, Crohn's Disease, or Ulcerative Colitis;

b) isolating and purifying from the T cell population a subpopulation of CD4+CD25+ Tregs; and

c) expanding the Treg cells of the subpopulation over 1,000 fold, in the presence of IL-2, anti-CD3 antibodies, and anti-CD28 antibodies, wherein the expanded Treg cells exhibit enhanced suppressive activity compared to a population of freshly purified, unexpanded Tregs from the individual;

wherein the anti-CD3 antibodies and anti-CD28 antibodies are immobilized on a three-dimensional solid surface.

2. The method of claim 1 , wherein the immune-mediated disease is Systemic Lupus Erythematous.

3. The method of claim 1 , wherein the immune-mediated disease is Multiple Sclerosis.

4. The method of claim 1 , wherein the immune-mediated disease is asthma.

5. The method of claim 1 , wherein the immune-mediated disease is Rheumatoid Arthritis.

6. The method of claim 1 , wherein the immune-mediated disease is Crohn's Disease.

7. The method of claim 1 , wherein the immune-mediated disease is Ulcerative Colitis.

8. The method of claim 1 , wherein the expansion is carried out over a period of about 3 weeks.

9. The method of claim 1 , wherein the three-dimensional solid surface is a bead.

10. The method of claim 9 , wherein a Treg to bead ratio is 1:3.

11. The method of claim 9 , the bead is about 1 to 10 microns in diameter.

12. The method of claim 9 , wherein an anti-CD3 and anti-CD28 antibody ratio on the surface of the bead is from about 1:20 to about 20:1.

Assignments (15)
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Mar 23, 2026
From: THERAKOS DEVELOPMENT LIMITED
To: THERAKOS HOLDINGS USA, LLC
Reel/Frame 075227/0886 →
PATENT SECURITY AGREEMENT Recorded Mar 17, 2026
From: THERAKOS HOLDINGS USA, LLC
To: NOMURA CORPORATE FUNDING AMERICAS, LLC, AS ADMINISTRATIVE AGENT AND COLLATERAL AGENT
Reel/Frame 075102/0334 →
RELEASE OF PATENT SECURITY INTEREST RECORDED AT REEL 69476, FRAME 0380 Recorded Mar 17, 2026
From: NOMURA CORPORATE FUNDING AMERICAS, LLC, AS ADMINISTRATIVE AGENT AND COLLATERAL AGENT
To: THERAKOS DEVELOPMENT LIMITED
Reel/Frame 075101/0970 →
RELEASE OF SECURITY INTEREST Recorded Aug 14, 2025
From: ACQUIOM AGENCY SERVICES LLC
To: MALLINCKRODT ENTERPRISES LLC; MALLINCKRODT LLC; MALLINCKRODT PHARMA IP TRADING UNLIMITED COMPANY; MALLINCKRODT PHARMACEUTICALS IRELAND LIMITED; OCERA THERAPEUTICS LLC; SPECGX LLC; STRATATECH CORPORATION; SUCAMPO PHARMA AMERICAS LLC; VTESSE LLC
Reel/Frame 072324/0740 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Mar 25, 2025
From: MALLINCKRODT PHARMACEUTICALS IRELAND LIMITED
To: THERAKOS DEVELOPMENT LIMITED
Reel/Frame 070624/0035 →
RELEASE OF SECURITY INTEREST Recorded Dec 3, 2024
From: ACQUIOM AGENCY SERVICES, LLC, AS COLLATERAL AGENT
To: MALLINCKRODT PHARMACEUTICALS IRELAND LIMITED
Reel/Frame 069463/0229 →
PATENT SECURITY AGREEMENT Recorded Dec 3, 2024
From: SOLARIS IPCO LIMITED
To: NOMURA CORPORATION FUNDING AMERICAS, LLC, AS ADMINISTRATIVE AGENT AND COLLATERAL AGENT
Reel/Frame 069476/0380 →
CHANGE OF NAME Recorded Nov 4, 2024
From: MALLINCKRODT HOSPITAL PRODUCTS IP LIMITED
To: MALLINCKRODT HOSPITAL PRODUCTS IP UNLIMITED COMPANY
Reel/Frame 069302/0292 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Nov 4, 2024
From: MALLINCKRODT HOSPITAL PRODUCTS IP UNLIMITED COMPANY
To: MALLINCKRODT PHARMACEUTICALS IRELAND LIMITED
Reel/Frame 069129/0753 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Nov 4, 2024
From: LI, LILY; CAO, TINGHUA
To: THERAKOS, INC.
Reel/Frame 069128/0830 →
NUNC PRO TUNC ASSIGNMENT Recorded Nov 4, 2024
From: THERAKOS, INC.
To: MALLINCKRODT CRITICAL CARE FINANCE INC.
Reel/Frame 069128/0976 →
NUNC PRO TUNC ASSIGNMENT Recorded Nov 4, 2024
From: MALLINCKRODT IP
To: MALLINCKRODT HOSPITAL PRODUCTS IP LIMITED
Reel/Frame 069129/0511 →
NUNC PRO TUNC ASSIGNMENT Recorded Nov 4, 2024
From: MALLINCKRODT PHARMA IP TRADING DAC
To: MALLINCKRODT IP
Reel/Frame 069129/0221 →
NUNC PRO TUNC ASSIGNMENT Recorded Nov 4, 2024
From: MALLINCKRODT CRITICAL CARE FINANCE INC.
To: MALLINCKRODT PHARMA IP TRADING DAC
Reel/Frame 069129/0071 →
SECURITY INTEREST Recorded Apr 24, 2024
From: MALLINCKRODT PHARMACEUTICALS IRELAND LIMITED; STRATATECH CORPORATION
To: ACQUIOM AGENCY SERVICES LLC
Reel/Frame 067215/0426 →
Continuity (6)
Continuation 17515985 · Nov 1, 2021
Division 15981332 · May 16, 2018
Continuation 15094467 · Apr 8, 2016
Continuation 12781451 · May 17, 2010
Provisional Application 61179165 · May 18, 2009
Related Publication 20240191193A1 · Jun 13, 2024
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