BIOCHEMISTRY, BIOMEDICINE & PHARMACEUTICS

Sarepta Therapeutics: SRP-9003 rAAVr74.MCK.SCGB Gene Therapy for Limb-Girdle Dystrophy Clinical Stage and Regulatory Timeline SRP-9003 is an investigational gene transfer therapy intended to deliver gene SGCB to muscle tissue for the targeted production of the beta-Sarcoglycan in the muscle. Sarepta is conducting phase 1/2 clinical study to determine safety, efficacy, and effective dose of SRP-9003 in patients with Limb-Girdle Muscular Dystrophy Type 2E with SGCB loss of function mutation. All patients have been enrolled in the study and administered with SRP-9003. The sponsors are awaiting two years of safety and efficacy endpoints data to initiate the pivotal trial. Viral Vector and Gene Construct:  AAVrh74.MHCK7.hSGCB- MHCK7 is a tissue-specific promoter that drives the expression in the muscle tissue. AAVrh74 is an adeno associated virus isolated from rhesus monkey and have high tropism for skeletal and cardiac muscle.  Clinical Phase: Sarepta is conducting a nonrando...

Phenylketonuria  Phenylketonuria (PKU) is a genetic disorder of liver metabolism that arises primarily from the loss of function mutation of PAH gene encoding the hepatic enzyme phenylalanine (Phe) hydroxylase (PAH). Enzyme PAH converts Phe to produce tyrosine (Tyr) and subsequently funneled into other metabolic pathways.   Lack of functional PAH results in hyperphenylalaninemia after dietary intake of protein, causing elevated systemic Phe levels. In high levels of Phe, patients show neurocognitive impairment and behavior problems. The classical PKU results in progressive, irreversible neurological impairment during infancy and early childhood, if left untreated. The characteristic features of the PKU include hypopigmentation in skin, hair, and eyes; eczema; growth retardation; and a mousy odor caused by the accumulation of phenylacetate and phenyllactate. Market Analysis of PKU The global phenylketonuria treatment market size was valued at USD 445.8 million in 2018 ...

Double-Stranded scAAV vectors (Next Generation Vectors) The double-stranded AAV genome is generated by mutating the terminal resolution site sequence on one side of the inverted terminal repeats (ITR), leading to the production of self-complementary AAV (scAAV). A genome <2.5 kilobases (kb; approximately half the size of the wild-type AAV genome) can therefore be packaged as a dimer in the AAV vector with the package size limit of 4.7kb. Advantages of scAAV vector design A single-stranded AAV viral vector is widely used for gene therapy. Upon transduction, the single-stranded DNA must be converted into a transcriptionally active double-stranded form, which is a crucial rate-limiting step in transgene expression from an rAAV vector. With a scAAV double-stranded vector design, the synthesis of the complementary strand is not required. Double-stranded scAAV is less prone to DNA degradation after viral transduction, thereby increasing the number of copies of stable episomes Doub...

Immune response associated adverse events and hypersensitivity reactions are the major safety concerns related to enzyme replacement therapies. The immunogenicity and anti-drug antibody generation against these therapeutic proteins are heterogeneous among the populations. Identify and characterizing individual factors causal for heterogeneities enable a better understanding and prediction of the immunogenicity (unwanted immune response) of therapeutic products. Among these individual factors, the characterization of residual enzyme/proteins or cross-reactive immunological material (CRIM) status, individual disease genotype may play a vital role in understanding immunogenicity and impact on safety and efficacy. Measurement of Residual enzyme activity For genetic disorders of enzyme deficiencies, enzyme activity is measured in the clinically suspected patients and diagnoses are made based on the decreased enzyme activity. The enzyme activity in affected subjects is usually less than...

Clinical Stage Therapeutic Area Parkinson Disease Parkinson disease is a progressive disorder of the nervous system. The disorder affects several regions of the brain, especially an area called the substantia nigra that controls balance and movement.  Treatment Approach: VY-AADC is a gene therapy for Parkinson’s disease that is being developed by Voyager Therapeutics. The treatment is aimed at increasing the patient’s ability to produce dopamine, a signaling molecule that helps regulate movement and emotional responses, which improves motor function. The targeted cells in the central nervous system (CNS) are long-lived, non-dividing neurons, therefore, treatments delivered in a single dose may generate long-lasting, or even lifelong, benefits. Clinical Stage: Phase 3/Pivitol  Platform: Adeno associated (AAV2) Viral Delivery of Genes into the target cells of the CNS VYGR Chart by TradingView

Clinical Trial Design RESTORE-1 (Phase 2 Clinical Study) (NCT03562494): Ongoing A randomized, double-blind, placebo-controlled trial evaluating the safety and efficacy of VY-AADC for the treatment of Parkinson’s disease in 75-100 patients (each trial) with motor fluctuations that are refractory to medical management.  Population: PD diagnosis > 4 yrs, are not responding adequately to oral medications, and have > 3 hours of OFF time. Randomized (1:1) to one-time administration of VY-AADC (for a total dose of up to 2.5×1012 vector genomes) or placebo surgery.  Experimental Treatment: VY-AADC02 (NBIb-1817) A single administration of up to 3.6 x 10^12 vector genomes (vg) of VY-AADC02 Adeno-associated viral vector serotype 2 encoding human aromatic L-amino acid decarboxylase (AAV2- hAADC) infusion into the brain NATIVE Primary Outcome Measures : Change in ON time without troublesome dyskinesia as recorded by participant in Parkinson's Disease (PD) diary [ Time F...

The Brilliance Phase 1/2 Clinical Trial Of AGN-151587 (EDIT-101) For The Treatment Of LCA10 Description This is an open-label, single ascending dose study of AGN-151587 (EDIT-101) in adult and pediatric (ie, ages 3 to 17) participants with LCA10-IVS26. Approximately 18 participants will be enrolled in up to 5 cohorts to evaluate up to 3 dose levels of AGN-151587 in this study. AGN-151587 is a novel gene editing product designed to eliminate the mutation on the CEP290 gene that results in the retinal degeneration that defines LCA10-IVS26. Single-dose of AGN-151587 administered by subretinal injection surgery Participants will receive a single dose of AGN-151587 administered via subretinal injection in the study eye. Up to 5 cohorts across 3 doses will be enrolled in this study. Primary Outcome Measure Frequency of Adverse Events related to AGN-151587  Number of participants experiencing procedural related adverse events  Incidence of dose limiting toxicities Seconda...

Gene Therapy Clinical Study in Adult PKU (pheNIX, Phase 1/2) Detailed Description:   This study will evaluate the safety and efficacy of HMI-102 gene therapy in adult subjects with PKU due to PAH deficiency. Subjects will receive a single dose of HMI-102 administered intravenously. Up to 3 dose levels of HMI-102 may be investigated in this study. At a given dose level, a minimum of 2 subjects will be enrolled and dosed. The dosing of the first two subjects will be staggered. Following the evaluation of data from the first 2 subjects in a cohort, a decision can be made to either escalate to the next dose level or expand the cohort at the selected dose level. If the cohort is expanded, additional subjects will be randomized to receive HMI-102 or a concurrent delayed treatment control arm. Subjects in the delayed treatment control will be eligible to receive HMI-102 after 24 weeks. Sponsor: Homology Medicine  Experiment Design: HMI-102 is an AAVHSC15 vector containi...

MiSeq System  Technical Specification The MiSeq System offers the first DNA-to-data sequencing platform, integrating cluster generation, amplification, sequencing, and data analysis into a single instrument. The MiSeq benchtop instrument utilizes a double-sided, single-lane flow cell and reagent cartridge supplied in kit form. Sequencing is performed by recording the synthesis of DNA strands in clusters of sample templates attached to the flow cell. The MiSeq System leverages Illumina sequencing by synthesis (SBS) technology, the most widely used, next-generation sequencing chemistry worldwide. With the power of next-generation sequencing (NGS) delivered in a compact footprint, the MiSeq System is the ideal platform for rapid and cost-effective genetic analysis. The combination of rapid library preparation and the MiSeq System delivers a simple, accelerated turnaround time. The library may be prepared in just 90 minutes with Nextera™XT library prep reagents. The autom...

AAV or transgene associated T-cell Response may increase cytotoxicity and loss of transgene expression. In the earliest AAV2 factor IX clinical trials, transient transgene expression was seen for up to eight weeks before loss in a subset of patients. Subsequently, it was determined to be an AAV capsid-specific CD8+ cytotoxic T cell response that likely led to the decline of factor IX expression. Therefore, regulatory agency require sponsor to monitor the T-cell response against AAV viral vectors and transgene and their impact on safety and efficacy in clinical subjects. Vector design and Administration to Clinical Subject of Phase 1a (nonrandomized control trial) for treatment of Duchenne Muscular Dystrophy The gene construct containing the MHCK7 promoter and micro-dystrophin transgene packaged into the AAVrh74 capsid were administered a single dose of 2.0 × 1014 vg/kg rAAVrh74.MHCK7.micro-dystrophin in approximately 10 mL/kg. This dose was infused through peripheral limb vein o...

Introduction of Giroctocogene Fitelparvovec Gene Therapy for Hemophilia A Giroctocogene Fitelparvovec is an investigational gene therapy product for the treatment of Hemophilia A co-developed by Sangamo/Pfizer. Giroctocogene fitelparvovec (SB-525) is a liver-tropic recombinant adeno-associated virus (rAAV6) vector carrying a B-domain–deleted F8 gene that is delivered through a single IV infusion. Sponsor: Sangamo Therapeutics/ Pfizer Stage-  Phase 3 Phase 2 - NCT03061201 Phase 3 - NCT04370054 (First Patient Dose)  Clinical Outcome  Efficacy: Efficacy was observed with significant reduction of annualized bleeding rate and usage of factor VIII infusion in phase 2 at the vector dose 3e13 vg/kg (viral genome/kg) upto 36 weeks after treatment. Safety: In phase 2 clinical studies, 4/5 subject at a vector dose of 3e13 vg/kg showed transient elevation of transaminases and required corticosteroid therapy. One treatment related SAE was observed.  ...

AAVrh74.MHCK7.micro-dystrophin (Serapta Therapeutics- SRP- 9001) Clinical Stage and Regulatory Timeline SRP-9001 is an investigational gene transfer therapy intended to deliver its micro-dystrophin-encoding gene to muscle tissue for the targeted production of the micro-dystrophin protein. U.S. Food and Drug Administration (FDA) has granted Orphan and Fast Track designation to SRP-9001 (AAVrh74.MHCK7.micro-dystrophin). Sarepta’s expectation to have results from Study 102 in early 2021; and potentially submit biologic licencing application to FDA in 2021. Viral Vector and Gene Construct:  AAVrh74.MHCK7.micro-dystrophin, MHCK7 is a tissue specific promoter that drive the expression in the muscle tissue. AAVrh74 is an adenoassociated virus isolated from rhesus monkey and have high tropism for skeletal and cardiac muscle.  Clinical Phase: Sarepta is currently conducting a Phas e 2 ( NCT03769116)  randomized, double-blind, placebo-controlled study to safety and effica...

AVV9-minidystrophin (Pfizer- PF-06939926) for Duchenne Muscular Dystrophy  Clinical Stage and Regulatory Timeline PF-06939926 is an investigational gene transfer therapy intended to deliver its mini-dystrophin-encoding gene to muscle tissue for the targeted production of the micro-dystrophin protein.(AAV9.Muscle-promoter.micro-dystrophin). Pfizer is initiating phase 3 pivotal trial to determine safety, efficacy of the PF-06939926 with 3e14 vg/kg dose. Viral Vector and Gene Construct AAV9.MuscleProtomter.mini-dystrophin (3.9Kb), The propriety muscle promoter drives the expression of the minidystrophin gene in the muscle tissue. AAV9 have shown to have high tropism for skeletal and cardiac muscle.  Clinical Phase Pfizer is currently conducting a Phas e 3 (NCT04281485) ra ndomized, double-blind, placebo-controlled study to safety and efficacy of the exogenous gene transfer in DMD subjects. The Phase 1b study conducted to determine the safety and tolerability of the AAV...

 AAV9- microdystrophin (Solid Biosciences- SGT-001 ) Clinical Stage and Regulatory Timeline SGT-001 is an investigational gene transfer therapy intended to deliver its micro-dystrophin-encoding gene to muscle tissue for the targeted production of the micro-dystrophin protein.(AAV9.CK.micro-dystrophin). Solid Biosciences is conducting phase 1/2 clinical study (IGNITE) to determine safety of SG-001 dose at 2e14 vg/kg dose. Viral Vector and Gene Construct AAV9.CK.micro-dystrophin  gene construct consists of muscle specific CK promoter and drives the expression of the minidystrophin gene in the muscle tissue. AAV9 have shown to have high tropism for skeletal and cardiac muscle.  Clinical Phase Solid Biosciences is currently conducting a Phas e 2  ( NCT03368742 ) nonra ndomized clinical study to evaluate safety of the exogenous gene transfer in DMD subjects. The immunofluorescence staining of the muscle biospy showed the successful transfer and expression of micro...

Overview of Clinical Stage Therapeutic Area Relapsed/Refractory B cell non-Hodkin Lymphoma: Refractory non-Hodgkin lymphoma (NHL) is NHL that has not responded to initial treatment. The refractory disease may be a disease that is getting worse or staying the same. Relapsed non-Hodgkin lymphoma (NHL) is NHL that responded to treatment but then returns. CD19 is the earliest differentiation antigen of the B lineage and is ubiquitously expressed on all types of B lymphocytes except plasma cells, thereby representing an attractive target for B-cell non-Hodgkin lymphomas (NHL) or leukemia of B-cell origin. Defect: Cancer/Lymphoma Treatment Approach: CB-010 is an off-the-shelf allogeneic anti-CD19, CAR-T cell therapy created using CRISPR/Cas9 technology for patients with relapsed/refractory B cell non-Hodgkin lymphoma (B-NHL). Clinical Stage: Phase 1/2 (IND cleared

 Clinical Stage Therapeutic Area Adult Phenylketonuria (PKU) Metachromatic Leukodystrophy (MLD) Adult Phenylketonuria (PKU) Phenylketonuria is a disorder of phenylalanine metabolism. In the patients with uncontrolled PKU, the blood Phenylalanine (Phe) level may rise above 1200 µM concentration. The clinical manifestations include growth failure, microcephaly, seizures, and intellectual impairment caused by the accumulation of toxic by-products of Phe. Gene Defect: Mutation in the PAH gene that encodes for enzyme phenylalanine hydroxylase that converts phenylalanine to tyrosine. PKU is inherited in an autosomal recessive manner Treatment Approach: HMI-102 is an investigational gene therapy designed to deliver the functional copy of the PAH gene to the adult patients using the AAVHSC vector. Specifically, administration of the AAVHSC-based gene editing constructs may result in the integration of the human PAH gene in human hepatocytes Clinical Stage: Phase 1/2 Metach...

Clinical Stage Therapeutic Area Laber Congenital Amaurosis Sickle Cell Disease Laber Congenital Amaurosis 10 Leber congenital amaurosis is an eye disorder that primarily affects the retina, which is the specialized tissue at the back of the eye that detects light and color. People with this disorder typically have severe visual impairment beginning in infancy. Gene Defect: Mutation in the CEP290 gene lead to degeneration in ocular photoreceptor cells that are critical for vision. The disease is inherited in an autosomal dominant pattern.  Treatment Approach: EDIT-101 is an investigational therapy ( in vivo ) designed to remove the CEP290 mutation and correct the disorder.   Clinical Stage: Early Clinical Stage (Click here for details) Sickle Cell Disease Sickle cell disease is a group of disorders that affects hemoglobin, the molecule in red blood cells that delivers oxygen to cells throughout the body. People with this disease have atypical hemoglob...

Fast Performance Liquid Chromatography (FPLC) & High-Performance Liquid Chromatography (HPLC) AKTA avant (GE Healthcare) Technical Specification ÄKTA avant is a preparative chromatography system intended for method and process development. ÄKTA avant systems have two versions with different flow rate/pressure designed for screening and method optimization, and scaling up to larger columns. Advantage of AKTA Avant ÄKTA avant together with UNICORN software provides a platform for efficient process development  Design of Experiments (DoE), an experimental design tool, provides time and cost savings by capturing more precise information in fewer experiments  Integrated fraction collector with cooling functionality protects purified samples Automatic on-line buffer preparation using BufferPro reduces the time required for buffer blending and manual titration, increasing your productivity  UNICORN software provides intuitive and flexible method creation, system contro...