Ignite Creation Date:
2024-05-06 @ 9:56 AM
Last Modification Date:
2024-10-26 @ 12:22 PM
Study NCT ID:
NCT03118128
Status:
COMPLETED
Last Update Posted:
2017-07-06
First Post:
2017-04-07
Brief Title:
Metformin Reduce the Relapse Rate on Patients With B-cell Precursor Ph Negative Acute Lymphoblastic Leukemia
Sponsor:
Hospital General de Mexico
Organization:
Hospital General de Mexico
Study Overview
Official Title:
Effect of the Addition of Metformin Hydrochloride on the Prognosis of Patients With B-cell Precursor Ph Negative Acute Lymphoblastic Leukemia With High Expression of ABCB1 Gene
Status:
COMPLETED
Status Verified Date:
2017-04
Last Known Status:
None
Delayed Posting:
No
If Stopped, Why?:
Not Stopped
Has Expanded Access:
False
If Expanded Access, NCT#:
N/A
Has Expanded Access, NCT# Status:
N/A
Acronym:
None
Brief Summary:
Metformins Antitumor activity were identified from differens diabetic patients trials mainly associated to its mechanism of action and protein - kinase AMPK AMP-activated protein kinase activation According to Cancer and Diabetes International Consensus from 2012 diabetes increases the risk for developping cancer and metformin has an protector effect against cancer cells and has an impact on overall survival
Chemotherapy drug resistance induces treatment fail in oncology Metformin increases AMPK levels blocks PI3K phosphatidylinositol 3- kinase AKT mTORmammailian Target of Rapamycin pathway but few evidence associated with drug resistance gene expression
This is an experimental one-center study that pretends to stablish the effect of adding metformin 850 mg PO three times a day over the multi-drug resistance gene expression ABCB1 in de novo Acute Lymphoblastic Leukemia in one 7-days cycle with prednisone as pre-treatment- and on the induction remission treatment
Chemotherapy drug resistance induces treatment fail in oncology Metformin increases AMPK levels blocks PI3K phosphatidylinositol 3- kinase AKT mTORmammailian Target of Rapamycin pathway but few evidence associated with drug resistance gene expression
This is an experimental one-center study that pretends to stablish the effect of adding metformin 850 mg PO three times a day over the multi-drug resistance gene expression ABCB1 in de novo Acute Lymphoblastic Leukemia in one 7-days cycle with prednisone as pre-treatment- and on the induction remission treatment
Detailed Description:
The MDR Multidrug-Resistance Genes are implicated on the resistance of several types of cancer The most important on leukemia are the ABCB1 and ABCG2- ABCB1 are implicated on resistance and severity on Acute Myeloid Leukemia and pediatric Acute Lymphoblastic Leukemia Those transporters use ATP for use Metformin decrease the intracellular ATP Adenosine triphosphate reserve the by the activation of AMPK Recently on MCF7-Adr Michigan Cancer Foundation 7 breast cancer cell line cancer cell line Metformin block the function of the P-glycoprotein by the inhibition of the Nuclear Factor -kappa-B
The clinical evidence at this moment is limited based on small clinical trials or observational studies This study tries to evaluate the effect of the addition of Metformin to a standard chemotherapy regimen on patients who express high levels of expression of mRNA messenger ribonucleic acid ABCB1
Experimental protocol
The patients will be stratified on three groups The high-expression low expression and absent gene expression according the level mRNA of ABCB1 at diagnostic The samples are obtained from mononuclear peripheral blood cells
Extraction of total RNA Total RNA was isolated by TRIzol Invtirogen Life Technologies according to the manufacturers recomendations described by Chomczynski and Sacchi
The concentration and purity of total RNA was determined in a UV -vis spectrophotometer Thermo Scientific Genesis 10S UV-vis The integrity of the genetic material was confirmed by 15 agarose gel electrophoresis at 70 V for 40 min The RNA was stored at -80C until needed
Synthesis of cDNA Fort he synthesis of c DNA complementary deoxyribonucleic acid the amount of RNA used was 2µg for a final volume of 20 µg The RNA was mixed with 1 µl oligonucleotide 12-18 INVITROGEN Carlsbad CA and 1 µl de dNTPs deoxynucleoside triphosphate 10mM Applied Biosystems Roche After the addition of 4 µl of Buffer 5 X Tris- HCl hydrochloric acid 250mM KCl potassium chloride 375 mM MgCl2 15mM 2 µl de DTT dithiothreitol 01M and wáter all the mix was incubated at at 37 for two minutes and 1 µl of MMLVMoloney Murine Leukemia Virus Reverse Transcriptase Reverse transcriptase- enzyme 200u INVITROGEN Carlsbad CA and incubated at 37C for 50 minutes
Real-time polymerase chain reaction qRT-PCR analysis The mRNA expression levels of the ABCB1 Hs01069047 and glyceraldehyde 3-phosphate dehydrogenase GAPDH Hs00985689 genes were measured using the TaqMan gene expression assay Applied Biosystems Foster City CA USA The GAPDH gene was used as an endogenous control and each sample was analyzed in triplicate The relative expression levels were calculated using the 2-ΔΔCtmethod with bone marrow as a calibrator The high and low expression cut-off points were determined by the mean values observed in healthy donors
Treatment protocol All the patients recieve an induction remission treatment with an initial pre-induction phase with steroids prednisone from day -7 to day -1 -7 -6 25mg -5 -4 50mg -3 -2 75mg and on day -1 100mg The induction remission treatment is based on a 28 day treatment with prednisone 60mgm2Vincristine 15mgm2 maxium 2mg and Daunorubicin 60mgm2 on days 1 8 and 15 If the patient archive a Complete Remission continues with a consolidation therapy with sequential blocks of treatment that includes Cytarabine Etoposide Methotrexate If the patient still on remission at the end of consolidation therapy continues on maintenance phase that includes 6-mercaptopurine and a weekly dose of methotrexate for around two years
Response At day 28 of the induction remission treatment according the result of bone marrow sample the patientes will be declared on remission less than 5 blast or refractory 5 blast cell If the patient present at any point of treatment an increase on bone marrow blast count the patient is considered on relapse
The clinical evidence at this moment is limited based on small clinical trials or observational studies This study tries to evaluate the effect of the addition of Metformin to a standard chemotherapy regimen on patients who express high levels of expression of mRNA messenger ribonucleic acid ABCB1
Experimental protocol
The patients will be stratified on three groups The high-expression low expression and absent gene expression according the level mRNA of ABCB1 at diagnostic The samples are obtained from mononuclear peripheral blood cells
Extraction of total RNA Total RNA was isolated by TRIzol Invtirogen Life Technologies according to the manufacturers recomendations described by Chomczynski and Sacchi
The concentration and purity of total RNA was determined in a UV -vis spectrophotometer Thermo Scientific Genesis 10S UV-vis The integrity of the genetic material was confirmed by 15 agarose gel electrophoresis at 70 V for 40 min The RNA was stored at -80C until needed
Synthesis of cDNA Fort he synthesis of c DNA complementary deoxyribonucleic acid the amount of RNA used was 2µg for a final volume of 20 µg The RNA was mixed with 1 µl oligonucleotide 12-18 INVITROGEN Carlsbad CA and 1 µl de dNTPs deoxynucleoside triphosphate 10mM Applied Biosystems Roche After the addition of 4 µl of Buffer 5 X Tris- HCl hydrochloric acid 250mM KCl potassium chloride 375 mM MgCl2 15mM 2 µl de DTT dithiothreitol 01M and wáter all the mix was incubated at at 37 for two minutes and 1 µl of MMLVMoloney Murine Leukemia Virus Reverse Transcriptase Reverse transcriptase- enzyme 200u INVITROGEN Carlsbad CA and incubated at 37C for 50 minutes
Real-time polymerase chain reaction qRT-PCR analysis The mRNA expression levels of the ABCB1 Hs01069047 and glyceraldehyde 3-phosphate dehydrogenase GAPDH Hs00985689 genes were measured using the TaqMan gene expression assay Applied Biosystems Foster City CA USA The GAPDH gene was used as an endogenous control and each sample was analyzed in triplicate The relative expression levels were calculated using the 2-ΔΔCtmethod with bone marrow as a calibrator The high and low expression cut-off points were determined by the mean values observed in healthy donors
Treatment protocol All the patients recieve an induction remission treatment with an initial pre-induction phase with steroids prednisone from day -7 to day -1 -7 -6 25mg -5 -4 50mg -3 -2 75mg and on day -1 100mg The induction remission treatment is based on a 28 day treatment with prednisone 60mgm2Vincristine 15mgm2 maxium 2mg and Daunorubicin 60mgm2 on days 1 8 and 15 If the patient archive a Complete Remission continues with a consolidation therapy with sequential blocks of treatment that includes Cytarabine Etoposide Methotrexate If the patient still on remission at the end of consolidation therapy continues on maintenance phase that includes 6-mercaptopurine and a weekly dose of methotrexate for around two years
Response At day 28 of the induction remission treatment according the result of bone marrow sample the patientes will be declared on remission less than 5 blast or refractory 5 blast cell If the patient present at any point of treatment an increase on bone marrow blast count the patient is considered on relapse
Study Oversight
Has Oversight DMC:
None
Is a FDA Regulated Drug?:
None
Is a FDA Regulated Device?:
None
Is an Unapproved Device?:
None
Is a PPSD?:
None
Is a US Export?:
None
Is an FDA AA801 Violation?:
None