CLINICAL TRIALS & FUNDING
Not long ago our good friend TT posted all the achievements of TSI without fundings. Around 2.5B shares were issued in about 10 years. I know of companies that just issued 2.8B shares in 3 months and are trading at $0.001, unlike TSI, which speaks of the great capabilities of our CEO Tim Dixon, Thomas, James and the BOD.
Let's see what 2.5B shares did for TSOI, this is without fundings...
65 filed patents
20 published patents
3 issued patents
(5) INDs w #’s
JadiCell/CTE Excl license
Treated (5) Nsvy SEALS successfully/CTE via RTT
International Amazon Prime sales
StemVacs Platform
Campbell Neurosciences
Allogen Biologics
CampbellCell
Campbell Score
ApoptoCyte
NanoStilbene
QuadraMune
Projuvenol
NanoCannabidiol
NanoPlus
NanoPSA
DermalStilbene
Now having $10M available, besides the ongoing Phase III clinical trial, do you know how many other potential clinical trials are waiting in line? And for a company to get a shot for a Clinical Trial, the inventors gotta be really really good at it, let's take a look of what could go thru clinical trials and how effective they are...
THERAPEUTIC MONOCYTES FOR PREVENTION OF SUICIDAL IDEATION
Type: Application
Filed: March 4, 2022
Publication date: September 8, 2022
Applicant: Therapeutic Solutions International, Inc.
Inventors: Thomas E. Ichim, Timothy G. Dixon, Famela Ramos, Kalina O'Connor, James Veltmeyer
Suppression of Inflammation Induced Memory Dysfunction by Cord Blood Derived Monocytes
The lipopolysaccharide administration model was used to replicate neuroinflammation associated with suicidal behavior. Female BALB/c mice were treated with control, daily lipopolysaccharide treatment (10 ng/mouse0, and some with cord blood derived monocytes. Cord blood derived monocytes were extracted by ficoll separation of cord blood mononuclear cells and 4-hour incubation on T175 plastic flasks. Non-adherent cells were washed off by rinsing with phosphate buffered saline and adherent cells where subsequently dissociated using trypsin and placed into another flask. Cord blood monocytes where cultured for 24 hours with 1 IU of oxytocin per million cells in complete DMEM media with 10% fetal calf serum. Cells were dissociated with trypsin, washed in phosphate buffered saline and administered intravenously vial tail vein at a concentration of 500,000 per mouse on days 0, 3, and 6.
To assess memory function, water filled basin which was 120 cm in diameter was broken into 4 quadrants. 10 cm diameter platform placed 1 cm below water. Mice were forced to swim to find the hidden platform, starting from all four different quadrants, each day for a total of 7 days. As seen below, mice receiving umbilical cord monocytes resisted the pathological effects of lipopolysaccharide treatment.
Suppression of LPS Induced Plasma Interleukin-6 by Cord Blood Derived Monocytes
The lipopolysaccharide administration model was used to replicate neuroinflammation associated with suicidal behavior. Female BALB/c mice were treated with control, daily lipopolysaccharide treatment (10 ng/mouse0, and some with cord blood derived monocytes. Cord blood derived monocytes were extracted by ficoll separation of cord blood mononuclear cells and 4-hour incubation on T175 plastic flasks. Non-adherent cells were washed off by rinsing with phosphate buffered saline and adherent cells where subsequently dissociated using trypsin and placed into another flask. Cord blood monocytes where cultured for 24 hours with 1 IU of oxytocin per million cells in complete DMEM media with 10% fetal calf serum. Cells were dissociated with trypsin, washed in phosphate buffered saline and administered intravenously vial tail vein at a concentration of 500,000 per mouse on days 0, 3, and 6.
To assess interleukin 6 levels in plasma, mice were sacrificed at the indicated timepoints and cytokine was assayed by ELISA.
EX VIVO GENERATION OF IMMUNOCYTES RECOGNIZING BROTHER OF THE REGULATOR OF IMPRINTED SITES (BORIS) EXPRESSING CANCER STEM CELLS.
Type: Application
Filed: February 22, 2022
Publication date: August 25, 2022
Applicant: Therapeutic Solutions International, Inc.
Inventors: Thomas E. Ichim, Timothy G. Dixon, Feng LIN, Famela Ramos, James Veltmeyer
EXAMPLES
Stimulation of Immunity to CD133 Positive PC-3 Cancer Stem Cells
Cord blood generated allogeneic dendritic cells were obtained by culture of adherent monocytes with GM-CSF (100 IU/ml) and IL-4 (100 IU/ml) for 7 days. Cells expressed CD11c and possessed typical dendritic morphology. Cells where incubated with full length BORIS protein and treated with 30 ng/ml Poly (IC) for 24 hours to induce dendritic cell maturation. Dendritic cells where cultured with allogeneic peripheral blood mononuclear cells for 7 days in the presence of IL-2 (10 IU/ml) and anti-CD3 and anti-CD28 beads (10(5) beads per ml. T cells where purified by Magnetic Activated Cell Sorting for CD3 (containing both CD8 and CD4 cells). PC-3 prostate cancer cells where cultured and separated into either control (no T cells added), unfractionated, CD133 negative and CD133 positive. Cells where co-cultured at a 1:1 ratio and cytotoxicity was determined by MTT assay.
As observed, T cells possessed a preferential ability to kill CD133+ cells, which corresponds to a cancer stem cell phenotype. Results are shown in FIG. 1.
Stimulation of Immunity to Fik Positive PC-3 Cancer Stem Cells
Cord blood generated allogeneic dendritic cells were obtained by culture of adherent monocytes with GM-CSF (100 IU/ml) and IL-4 (100 IU/ml) for 7 days. Cells expressed CD11c and possessed typical dendritic morphology. Cells where incubated with full length BORIS protein and treated with 30 ng/ml Poly (IC) for 24 hours to induce dendritic cell maturation. Dendritic cells where cultured with allogeneic peripheral blood mononuclear cells for 7 days in the presence of IL-2 (10 IU/ml) and anti-CD3 and anti-CD28 beads (10(5) beads per ml. T cells where purified by Magnetic Activated Cell Sorting for CD3 (containing both CD8 and CD4 cells). PC-3 prostate cancer cells where cultured and separated into either control (no T cells added), unfractionated, flk negative and flk positive. Cells where co-cultured at a 1:1 ratio and cytotoxicity was determined by MTT assay.
As observed, T cells possessed a preferential ability to kill PC-3 flk+ cells, which corresponds to a cancer stem cell phenotype. Results are shown in FIG. 2.
Stimulation of Immunity to Rhodamine Positive PC-3 Cancer Stem Cells
Cord blood generated allogeneic dendritic cells were obtained by culture of adherent monocytes with GM-CSF (100 IU/ml) and IL-4 (100 IU/ml) for 7 days. Cells expressed CD11c and possessed typical dendritic morphology. Cells where incubated with full length BORIS protein and treated with 30 ng/ml Poly (IC) for 24 hours to induce dendritic cell maturation. Dendritic cells where cultured with allogeneic peripheral blood mononuclear cells for 7 days in the presence of IL-2 (10 IU/ml) and anti-CD3 and anti-CD28 beads (10(5) beads per ml. T cells where purified by Magnetic Activated Cell Sorting for CD3 (containing both CD8 and CD4 cells). PC-3 prostate cancer cells where cultured and separated into either control (no T cells added), unfractionated, Rhodamin negative and Rhodamin positive. Cells where co-cultured at a 1:1 ratio and cytotoxicity was determined by MTT assay.
As observed, T cells possessed a preferential ability to kill PC-3 Rhodamin+ cells, which corresponds to a cancer stem cell phenotype.
STIMULATION OF NATURAL KILL CELL MEMORY BY ADMINISTRATION OF DENDRITIC CELLS
Type: Application
Filed: February 8, 2022
Publication date: August 11, 2022
Applicant: Therapeutic Solutions International, Inc.
Inventors: Thomas E. ICHIM, Timothy G. DIXON, James VELTMEYER, Famela RAMOS
Example 1 Homotaurine Increases Ability of StemVacs to Stimulate NK Activity
Peripheral blood mononuclear cells where obtained by ficoll centrifugation and plated with StemVacs umbilical cord derived dendritic cells. StemVacs was generated by culture of umbilical blood adherent cells with interleukin 4 and GM-CSF for 7 days with maturation step induced by 24 hour culture TLR agonist. Assessment of natural killer cell activity was performed subsequent to culture of cells with taurine (10 micrograms/ml) or homotaurine (10 micrograms/ml) for the indicated timepoints. Cytotoxicity against NK target cell line was performed using the flow cytometry Promega assay. Results are shown in FIG. 1.
Example 2 Homotaurine Increases Ability of StemVacs to Stimulate T Cell Activity
Peripheral blood mononuclear cells where obtained by ficoll centrifugation and plated with StemVacs umbilical cord derived dendritic cells. StemVacs was generated by culture of umbilical blood adherent cells with interleukin 4 and GM-CSF for 7 days with maturation step induced by 24 hour culture TLR agonist. Assessment of T cell activity was performed subsequent to culture of cells with taurine (10 micrograms/ml) or homotaurine (10 micrograms/ml) for the indicated timepoints. T cell activity was assessed by ELISA quantification of interferon gamma production after stimulation with 5 micrograms per ml of phytohemagglutinin.
IMMUNOTHERAPY FOR OPIOID ADDICTION
Type: Application
Filed: December 20, 2021
Publication date: June 23, 2022
Applicant: Therapeutic Solutions International, Inc.
Inventors: Thomas Ichim, Timothy G. Dixon, Famela Ramos, Wais Kaihani, James Veltmeyer, Kalina O'Connor
Example 1: Reduction of Brain Microglial Activation by PRP and Pterostilbene
BALB/c mice were anesthetized with isofluorane and administered pterstilbene (0.4 mg/mouse) and/or platelet rich plasma (5 microliter per mouse) via intranasal route subsequent to induction of systemic inflammation by intraperitoneal administration of endotoxin. Mice were sacrificed after 24 hours of treatment and quantification of cytokine IL-18 was performed by ELISA from brain homogenate tissue. As seen below, a synergistic reduction of TNF-alpha was observed. Results are shown in FIG. 1.
Example 2: Reduction of Brain Microglial Activation by PRP and Oxytocin
BALB/c mice were anesthetized with isofluorane and administered oxytocin (0.1 IU/mouse) and/or platelet rich plasma (5 microliter per mouse) via intranasal route subsequent to induction of systemic inflammation by intraperitoneal administration of endotoxin. Mice were sacrificed after 24 hours of treatment and quantification of cytokine IL-18 was performed by ELISA from brain homogenate tissue. As seen below, a synergistic reduction of TNF-alpha was observed. Results are shown in FIG. 2.
TREATMENT OF MAJOR DEPRESSIVE DISORDER AND SUICIDAL IDEATIONS THROUGH STIMULATION OF HIPPOCAMPAL NEUROGENESIS UTILIZING PLANT-BASED APPROACHES
Type: Application
Filed: December 8, 2021
Publication date: June 9, 2022
Applicant: THERAPEUTIC SOLUTIONS INTERNATIONAL, INC.
Inventors: Thomas Ichim, Timothy G. Dixon, James Veltmeyer, Kalina O'Connor
Example
BALB/c mice where exposed to random stress 4 times a day by spinning by the tail for 15 seconds. Controls where not stressed. Stressed mice where treated with Prozac or QuadraMune™ daily. Assessment of endogenous neurogenesis was performed by administering BRDU and assessment of incorporation by histology. Augmented neurogenesis was seen in animals which received Prozac, with enhanced neurogenesis in animals taking QuadraMune™
PROTECTION AND REGENERATION OF NEUROLOGICAL FUNCTION BY USING STEM CELLS
Type: Application
Filed: October 27, 2021
Publication date: April 28, 2022
Applicant: Therapeutic Solutions International, Inc.
Inventors: Thomas E. Ichim, Timothy G. Dixon, Amit N. Patel, Famela Ramos, Kalina O'Connor
Examples
JadiCell Conditioned Media Inhibits Endothelial Death.
Injury associated with inflammation. Inflammation causes oxidative stress. Oxidative stress induces death of endothelial cells. Endothelial death exposes basement membrane, induces coagulopathy. Culture of HUVEC cells with upstream inflammatory agent TNF-alpha or downstream H2O2 results in death. Conditioned media (CM) from JadiCells decreases endothelial cell death. Results are shown in FIGS. 1 and 2.
JadiCell™ Reduces Immunogenicity of Endothelial Cells
Endothelial cells can act as antigen presenting cells. Stimulation of T cells by endothelial cells results in inflammation and breaking of blood brain barrier. HLA expression on HUVEC was utilized to quantify one aspects of endothelial antigen presentation. Mixed lymphocyte reaction used as a test of T cell activation. Results are shown in FIGS. 3 and 4.
JadiCell™ Reduces Thrombogenicity of Activated Endothelial Cells
In response to inflammation endothelial cells induce clotting by stimulation of extrinsic coagulation pathway. Tissue factor is activator of extrinsic pathway. HUVEC cells were treated with endotoxin to stimulate activation. Assessment of Tissue Factor performed by flow cytometry. Results are shown in FIG. 5.
CELLULAR, ORGAN, AND WHOLE-BODY REJUVENATION UTILIZING CORD BLOOD PLASMA AND PTEROSTILBENE
Type: Application
Filed: November 4, 2020
Publication date: May 6, 2021
Applicant: Therapeutic Solutions International, Inc.
Inventors: Thomas E. Ichim, Timothy G. Dixon
EXAMPLE
Cord Blood Plasma Suppression of Senescence Marker Beta Galactosidase is Augmented by Pterostilbene
Foreskin fibroblasts where obtained from ATCC and cultured according to manufacture instructions. Accelerated senescence was induced by exposure to indicated concentrations of H2O2 for 48 hours. Cells where cultured in control media (DMEM) or 10% cord blood plasma, or combination of cord blood plasma with 3 uMol Pterostilbene. Senescence was detected by fixing cells with 4% paraformaldehyde and SA-ß-Gal was stained using senescent cells histochemical staining kit (Sigma Aldrich, St. Louis, Mo., USA). Three images per each well were collected, and the SA-ß-Gal-stained cells were counted.
THIS IS JUST TO SHOW A FEW OF THE EXPERIMENTS AND RESULTS SEEN ON THE EXAMPLES, THOUGH TSOI ALREADY HAD SUCCESSFUL TRIALS IN THE PAST, WITH THIS REPUTATION,.I HAVE NO DOUBTS THAT TSOI IS GOING TO SHAKE THE WHOLE BIOPHARMA INDUSTRY.
