CA-3152963-C - ACTIVATED MESENCHYMAL STEM CELLS FOR TREATING LIMB ISCHEMIA

Abstract

The invention provides metamizole treated mesenchymal stem cells for treating limb ischemia of a human patient. The invention provides a pharmaceutical product for angiogenesis growth to replace occluded blood vessels for avoiding limb amputation. Secretion of angiogenic growth factors VEGFA, HGF, bFGF, TEK are stimulated and the levels of pro-inflammatory cytokines IF6, CXCF8, CCF2, IL1-RN are reduced by activation of mesenchymal stem cells (MSCs). According to treatment results, MSCs produce proteins and signalling molecules for new blood vessel growth that accelerate the growth of new arteries.

Inventors

• Andrus LOOG
• Jekaterina Kazantseva
• Olavi VASAR
• Tiit MEREN
• Triin VASAR
• Mart RAIK

Assignees

• CELLIN TECHNOLOGIES OU
• TAASTAVA KIRURGIA KLIINIK AS

Dates

Publication Date

20260505

Application Date

20190827

Claims (2)

1. CLAIMS: 1. Product comprising metamizole treated mesenchymal stem cells for use in the treatment of human limb ischemia.
2. 2. An in vitro use of a metamizole for activation of mesenchymal stem cells (MSCs) for stimulating secretion of angiogenic growth factors VEGFA, HGF, bFGF, TEK and reducing the levels of pro-inflammatory cytokines IL6, CXCLB, CCL2, IL 1- RN. Date Re9ue/Date Received 2023-11-22

Description

1 Activated mesenchymal stem cells for treating limb ischemia Field of the Invention Progress and success of the cell therapy is dependent of technologies that enable us to manipulate stem cells by stimulation of proliferation, differentiation and integration into 5 regenerating tissues and organs. Isolated stem cells can be manipulated at different stages of cell therapy procedures including cell isolation, cell propagation, conditioning for transplantation and post grafting. The present invention relates to the use of clinically approved anti-inflammatory drugs to modulate activity of mesenchymal stem cells isolated from the adipose tissue. Modulation of secretion of different growth factors including angiogenic factors 10 and cytokines as well as modulation of metabolic activity can be used to develop efficient MSC based pharmaceutical products for treatment of variety of health conditions. Background Mesenchymal stem cells (MSCs) are characterized by their ability to differentiate into variety of cell lineages in vitro and to have immunomodulatory function in regenerative processes 15 (Augello & De Bari, 2010), (Shi et al., 2010). Unlike pharmaceutical treatments that deliver particular active component at a specific dose, MSCs exert therapeutic effects by secreting various bioactive compounds in response to external stimulation (Ma et al., 2014). The soluble factors produced by MSCs are involved in anti-inflammatory and neovascularisation processes with profound effects on tissue injury and regeneration. Among diseases that can be treated 20 using MSCs are for example immune and non-immune disorders such as myocardial infarction, diabetes, graft versus host disease, and liver cirrhosis (Wei et al., 2013) (Shi et al., 2010). Understanding the immunomodulatory properties of MSCs and ways how inflammatory microenvironment affects their function is of immense importance for developing better strategies to increase therapeutic efficiency of MSCs with a goal to create local and/or systemic 25 conditions to stimulate healing and tissue regeneration. Peripheral arterial disease is a condition characterized by restricted supply of oxygen and glucose due to malfunction of blood vessels. Peripheral vascular disease commonly affects arteries and in most advanced stages causes critical limb ischemia (CLI). Up to date, the most 2 common therapeutic options include pharmacological treatments and surgery (Norgren et al., 2007), (Hamdy et al., 2013), (Cull et al., 2014). Non-steroidal anti-inflammatory drugs and synthetic glucocorticoid injections are widely used to reduce pain and inflammation of CLI patients. However, in many cases the result of the disease progression is gangrene and limb 5 amputation. Development of new advanced therapies could improve clinical outcome and increase the life standard of patients. The clinical potential of MSCs for the treatment of ischemic conditions has been described in several animal models and early-phase human clinical trials (Liew & O'Brien, 2012). The efficiency and safety of administration of MSCs for treatment of acute ischemic disorders show great potential. Therapeutic efficiency of MSCs 10 depends on their ability to provide immunomodulatory and angiogenic factors to suppress inflammation and stimulate angiogenesis. Non-activated MSCs express low levels of immunosuppressive factors, but the local conditions at the site of injections affect their functionality. To improve the therapeutic effect of MSCs, different strategies have been developed. The stimulation of MSCs by IFNy or TNFa has been used to induce the secretion 15 of immunomodulatory factors (Crop et al., 2010). Also, overexpression of CXCR4 in MSCs results in more effective homing of MSCs into ischemic tissue compared to unmodified cells (Cheng et al., 2008). Thus, immunosuppressive and angiogenic effects of MSCs could be stimulated by changing the conditions in the affected/diseased tissue or pretreatment of MSCs prior the grafting. Since patients with limb ischemia are treated with different anti-inflammatory 20 drugs, the understanding of the consequences of these drugs on the anti-inflammatory and angiogenic function ofMSCs is extremely important. The majority ofNSAIDs function through the blockade of prostaglandin synthesis by inhibition of cyclooxygenase enzyme. Prostaglandin PGE2 is known to be one of the important compounds secreted by MSCs that is responsible for modulation of inflammation. However, the effect of NSAIDs on complex of metabolic 25 responses and secretion of anti-inflammatory and angiogenic factors in MSC anti-inflammatory therapy is not known. The effect of NSAIDs (paracetamol, metamizole (analgin), ketoprofen and diclofenac) and glucocorticoid prednisolone, used in clinical practice to treat ischemic disorders, at therapeutic doses on cell cycle, metabolic activity, as well as on expression of angiogenic and inflammatory 30 cytokines by AdMSCs has been analysed and will be described he