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LYSINE BIOTECH PRIVATE LIMITED
 Product development and Manufacture of drugs, biomarkers and vaccines from Protein and DNA
 CIN:U74999TN2018PTC122879
 

Research and Development

To develop low cost and affordable IND/Biobetter drug for treating Type 2 diabetes-ilirapeptide/ilirag1 (GLP-1)To develop low cost and affordable IND/Biobetter drug for treating Type 2 diabetes-ilirapeptide/ilirag1

The goal of objective:

In this project proposal is to discover and develop the nature derived conventional solid phase peptide synthesis of drug for type 2 diabetes and other related diseases using an in vivo mouse model and human clinical trials. The new strategy and methodology of unique peptide synthesis apply to solve the problems of biological and medical importance and ultimately impact human health.

Background:

For the past one decade, peptide therapeutics playing a significant role in drug discovery. Consequently, there is an increased interest on peptides in pharmaceutical research and development. There are approximately 360 peptide drugs are in clinical trials and 8 drugs are recently approved by US FDA for treatment of Type-2 Diabetes (Liraglutide and semeglutide) and other diseases. Many of these clinically approved peptides based drugs shows promising biological activity and entered into human clinical trials and very few of them have been approved by US FDA.

Process:

Solid-phase peptide synthesis relies on assembling peptide chain on an insoluble polymeric resin support, thereby simplifying the process.

Deliverable:

Since existing peptides has limited synthetic routes with vast limitations, it has now become necessary to explore novel approaches, for synthesis of existing/new peptides that are beyond traditional solid phase peptide synthesis is very economical method for the synthesis amino acid peptides that result in high purities. Production cost less expensive due to the convergent approaches.

Target beneficiary and impact:
Drawbacks in the existing state of art Novo Nordisk.

Liraglutide and semeglutide are a fragment of the naturally occurring human glucagon-like peptide-1 sequence position 7-37 (GLP-1) and with addition of a fatty acid chain (palmitoyl group on Lys). liraglutide and semeglutide higher cost drug protocol higher dosage from protein chemical formula C172H265N43O51 and C187H291N45O59 Protein average weight 3751.2 Da and 4113.64 Da on the other hand, long peptide-based drugs generally suffer from short half-lives due to proteolytic degradation and fast renal clearance, which require high cost and higher doses/frequency of injections liraglutide 0.6 mg, for once- 1 day and semeglutide 0.25 mg, for once-7 days.

Side effect related to thyroid cancer and other diseases.

How these drawbacks have been overcome by LYSINE BIOTECH

ilirapeptide/ilirag1 IND/Biobetter drug is a fragment of the naturally occurring human glucagon-like peptide-1 sequence position 7-37 (GLP-1). low cost effective drug ilirapeptide/ilirag1 lower dosage from protein molecular formula C60H89N23O19 and Protein average weight 1435 Da on the other hand, low cost and lower doses of injections ilirapeptide/ilirag1 0.2 mg, for once-8 days

No side effect related to any other diseases.

This research is of utmost importance for the treatment of type 2 diabetes and other incertin related diseases.

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Human coronavirus hCoV-19/India 1st, 2nd & 3rd waves COVID-19

Available.

We put emphasize that with this computationally developed, predicts protein sequence while the invention of new novel study suggest that thymine content (uracil) in protein coding RNA sequences are randomly distributed. Thymine content distribution is used here to examine hCoV-19/India genome (hCoV-19/ India types 1-3). Frames are arriving as expected, single-stranded RNA first three frames, in the hCoV-19/India genomes types 1-3. SPIKE & Nucleocapsid -Frame-1 and RdRp-Frame-3 prefer to have definite amount of protein coding hCoV-19/India -3rd COVID-19 wave 2021). SPIKE & Nucleocapsid -Frame-2 and RdRp-Frame-1 were observed the protein coding hCoV-19/India -2nd COVID-19 wave 2021 and SPIKE & Nucleocapsid -Frame-3 and RdRp-Frame-2 were also involved protein coding hCoV-19/India-1st COVID-19 wave 2020. In addition, the results reveal that there were difference between (hCoV-19/India-1st, 2nd & 3rd COVID-19 types 1-3.

HCoV-19/India-1st, 2nd & 3rd waves COVID-19 types 1-3, determined the role of thymine protein coding frames of RNA sequences (Bioinformation. 2008. 2, 304-307). In this results Human coronavirus (hCoV-19/India -1st, 2nd & 3rd waves COVID-19 types 1-3 protein coding RNA of Spike, Nucleocapsid and RdRp play an important role in such a way to establish host-pathogen interaction.

Severe acute respiratory syndrome-coronavirus-2 isolate Wuhan-Hu-1 SPIKE & Nucleocapsid-Frame-2 and RdRP-Frame-1: 1st COVID-19 wave 2020: 29903 bp +ssRNA
Human coronavirus hCoV-19/India/TG-RFCH00152_CIB1129/2020:SPIKE & Nucleocapsid-Frame-3 and RdRp-Frame-2: 1st COVID-19 wave 2020: 29814 bp RNA
Human coronavirus hCoV-19/India/TG-RFCH00254_CIB0664/2021:SPIKE & Nucleocapsid-Frame-2 and RdRp-Frame-1: 2nd COVID-19 wave 2021: 29014 bp RNA
Human coronavirus hCoV-19/India/TG-RFCH00327_CIB0737/2021:SPIKE & Nucleocapsid-Frame-1 and RdRp-Frame-3: 3rd COVID-19 wave 2021: 29784 bp RNA
2nd COVID-19 wave lockdown did not follow up meanwhile Severe acute respiratory syndrome-coronavirus-2 (SARS COV-2) affected people's died in India 2021
examle: Severe acute respiratory syndrome-coronavirus-2 isolate Wuhan-Hu-1 SPIKE & Nucleocapsid-Frame-2 and RdRP-Frame-1: 1st COVID-19 wave 2020: 29903 bp +ssRNA China.
Human coronavirus hCoV-19/India/TG-RFCH00254_CIB0664/2021: SPIKE & Nucleocapsid-Frame-2 and RdRp-Frame-1: 2nd COVID-19 wave 2021: 29014 bp RNA India.

Current State-Of-The-Art Problem

New insight into the various types of unique specific coronavirus (Alpha, Beta, Gamma, Delta, lamda & Omicron). In this study suggest that protein coding +ssRNA of Spike, Nucleocapsid, RdRp, NSP7-RdRp, NSP8-RdRp, NSP12-RdRp sequences in SARS-COV-2 are randomly trimer proteins distributed and used here to examine SARS-COVID-19 type 2. ORFs are arriving as expected, ssRNA positive-strand first three ORF 1,2 & 3 in the SARS-COV-2 virus genome. In this present study suggest that i)spike protein18 amino acids was man-made in lab to increased length of artificial gene/protein and changed the functional site position of N-linked and introduced newly two O-linked (PTM) SARS-COV-2 from SARS-COV-1. ii-iii) Nucleocapsid and RdRp protein 3aa & 1aa was man-made in lab to reduced length not artificial gene/protein SARS-C0V-2 from SARS-CoV-1. Bio computational study suggest that Enterobacteria phage T4 18bp/6aa available in the Spike 1273 SARS CoV-2. In this results SARS-COV-2 protein coding +ssRNA of RdRp, Spike and Nucleocapsid play an important role in such a way to establish host-pathogen interaction.

RBD-protein sub-unit vaccine developer Biological E/BCM, Anhui Zhifei Longcom Biopharmaceutical/Institute of Microbiology, Chinese Academy of Sciences and West China Hospital, Sichuan University P

Proposed Solution-COVID-19 & OMICRON by LYSINE BIOTECH

SASR-COV-2 (COVID-19) SPIKE-RBD,S1 & S2 etc., and Nucleocapsids sequence not patented are open sourced from LYSINE BIOTECH May 21, 2020

SASR-COV-2 (COVID-19) SPIKE-RBD-S1 & S2 protein sub-unit vaccine for 5 Age to 15 Age children for covid-19 developer LYSINE BIOTECH

Specific Goals SARS-COV-2 (COVID-19) by LYSINE BIOTECH

I) Development of Alpha, Beta, Gamma, Delta, lamda & Omicron specific RBD-S1, S2 and Nucleocapsid custom antibody detection kit using recombinant antigens. Set-up of antibody and antigen Rapid test kits prototype COVID-19.

II) Development of Alpha, Beta, Gamma, Delta, lamda & Omicron specific unique peptide anti-RBD SPIKE antiviral-treatment of COVID-19.

Proposal Application ID - 105 "Production of bispecific antiviral antibodies Receptor Binding Domain SARS COVID-19" for funding - NIDHI4COVID2.0
Application could not find place in the short listed startups selected for funding under NIDHI4COVID 2.0 (Aug 21, 2021)

III) Development of Alpha, Beta, Gamma, Delta, lamda & Omicron recombinant protein subunit vaccine RBD-S1 COVID-19.

Protein sequence target specific (N-terminal and C-terminal) based on antibody epitope prediction using Alpha, Beta, Gamma, Delta, lamda & Omicron for rapid test antigen and antibody to peptide/protein drugs up to vaccines for COVID-19
Recombinant Protein Expression and Purification of 524aa-575aa and 868aa-967aa RBD-S1 & S2 SPIKE and 52aa-195aa Nucleocapsid N protein for rapid test antigen and antibody. Set-up of antibody and antigen Rapid test kits prototype COVID-19
Recombinant Protein Expression and Purification of 524aa-575aa RBD-S1 protein sub-unit vaccine candidates for COVID-19 Platform/technology from Yeast, CHO Cells and Insect Cells
Recombinant Protein Expression and Purification of 524aa-575aa RBD-S1 & 868aa-967aa S2 SPIKE protein sub-unit vaccine candidates for COVID-19 Platform/technology from Yeast, CHO Cells and Insect Cells
Chemical synthesis and peptides drug and vaccines are usually composed of 7–44 amino acids containing the specific epitope of an antigen for COVID-19

Severe acute respiratory syndrome coronavirus 2 isolate Wuhan-Hu-1, complete genome COVID-19

>RBD-SPIKE-DELTA: VCGPKKSTNLVKNKCVNFNFNGLTGTGVLTESNKKFLPFQQFGRDIADTTDA

Two specific Epitopes DELTA

KKSTNLVKNKC
FNGLTGTGVLTESNKKFLPFQQFGRDIAD
>SPIKE-S2-DELTA: EMIAQYTSALLAGTITSGWTFGAGAALQIPFAMQMAYRFNGIGVTQNVLYENQKLIANQFNSAIGKIQDSLSSTASALGKLQDVVNQNAQALNTLVKQLS

One specific Epitope DELTA

GWTFGAGA
> Nucleocapsids-DELTA: WFTALTQHGKEDLKFPRGQGVPINTNSSPDDQIGYYRRATRRIRGGDGKMKDLSPRWYFYYLGTGPEAGLPYGANKDGIIWVATEGALNTPKDHIGTRNPANNAAIVLQLPQGTTLPKGFYAEGSRGGSQASSRSSSRSRNSSR

Three specific Epitopes DELTA

LTQHGKEDLKFPRGQGVPINTNSSPDDQIGYYRRATRRIRGGDGKMKDLSP
EAGLPYGANKD
GALNTPKDHIGTRNPANNAAIVL

OMICRON: SPIKE-RBD,S1 & S2 etc., and Nucleocapsids OMICRON sequence not patented are open sourced from LYSINE BIOTECH 29-11-2021

OMICRON from DELTA (SARS-COV-2) ONE Mutation in the RBD and S2 Spike Protein sequence of OMICRON

OMICRON from DELTA (SARS-COV-2) NO Muatation in the Nucleocaspid protein sequence of OMICRON

>hCoV-19/USA/MD-HP20874-PIDUYWZOWA/2021|EPI_ISL_7160424| OMICRON
>RBD-SPIKE-OMICRON: VCGPKKSTNLVKNKCVNFNFNGLKGTGVLTESNKKFLPFQQFGRDIADTTDA

One specific Epitopes OMICRON

KKSTNLVKNKCVNFNFNGLKGTGVLTESNKKFLPFQQFGRDIAD
>hCoV-19/USA/MD-HP20874-PIDUYWZOWA/2021|EPI_ISL_7160424|-OMICRON
>SPIKE-S2-OMICRON: MIAQYTSALLAGTITSGWTFGAGAALQIPFAMQMAYRFNGIGVTQNVLYENQKLIANQFNSAIGKIQDSLSSTASALGKLQDVVNHNAQALNTLVKQLSS

One Epitopes OMICRON

Q
>hCoV-19/USA/MD-HP20874-PIDUYWZOWA/2021|EPI_ISL_7160424|-OMICRON
>Nucleocapsids-OMICRON: WFTALTQHGKEDLKFPRGQGVPINTNSSPDDQIGYYRRATRRIRGGDGKMKDLSPRWYFYYLGTGPEAGLPYGANKDGIIWVATEGALNTPKDHIGTRNPANNAAIVLQLPQGTTLPKGFYAEGSRGGSQASSRSSSRSRNSSR

Three specific Epitopes OMICRON

LTQHGKEDLKFPRGQGVPINTNSSPDDQIGYYRRATRRIRGGDGKMKDLSP
EAGLPYGANKD
GALNTPKDHIGTRNPANNAAIVL

Proposed Solution-PANCOVID: Recombinant unique antigen Peptide Vaccine and RNA-Targeted small antiviral peptide Drug Discovery and development

Our computational studies provide new insight into the various types of unique specific coronavirus Alpha, Beta, Gamma, Delta, lamda & Omicron i) Retrieving all coronavirus protein IDs (protein coding ssRNA of Spike, Nucleocapsid, RdRp, NSP7-RdRp, NSP8-RdRp, NSP12-RdRp etc.,) from NCBI. ii) Checking the protein expression of all coronavirus protien across various types of coronavirus at the NCBI Virus sequences for discovery data base.iii) Calculating and visualization the protein expression correlation of top pancoronavirus protein candidates across various types of pancoronavirus using our in-house software analysis tools. iv) Coronavirus disease and disease -specific GeneX candidate identification: e.g.( i)fourteen in one Unique peptide ii)fourteen in one Unique peptide and iii)fifteen-in-One Unique peptide v) Development of GeneX viral-based Recombinant unique peptide vaccine and RNA-Targeted small unique antipeptide viral molecule Drug Discovery and development therapeutic candidates demonstrating potential efficacy against for PANCOVID

Specific Goals PANCOVID

i)fourteen-in-One, ii)fourteen-in-One and iii)fifteen-in-One GeneX viral-based Recombinant unique antigen peptide vaccine candidates demonstrating potential efficacy against PANCOVID

I)Development of Alpha, Beta, Gamma, Delta, lamda & Omicron 1st Fourteen-in-One GeneX viral-based Recombinant Unique antigen peptide vaccine for PANCOVID.

ICMR: Concept Proposal id 2021-5160 with title Development of subunit pan coronavirus vaccine for multiple microorganism of COVID
ICMR: Review by external expert, the council does not find it acceptable for further processing and expanding in the form of detailed proposal and hence term it 'Not Recommended (Jul 30, 2021)

II)Development of Alpha, Beta, Gamma, Delta, lamda & Omicron 2nd Fourteen-in-One GeneX viral-based Recombinant Unique antigen peptide vaccine for PANCOVID.

III)Development of Alpha, Beta, Gamma, Delta, lamda & Omicron 3rd Fifteen-in-One GeneX viral-based Recombinant Unique antigen peptide vaccine for PANCOVID.

i)fourteen-in-One, ii)fourteen-in-One and iii)fifteen-in-One tri-specific GeneX viral antiviral peptide Drug candidates with strong binding targeting multiple microorganisms on the PANCOVID

I)Development of Alpha, Beta, Gamma, Delta, lamda & Omicron 1st Fourteen-in-One GeneX viral-based RNA-Targeted small Unique antiviral peptide Drug to treat severe cases of PAN-COVID.

II)Development of Alpha, Beta, Gamma, Delta, lamda & Omicron 2nd Fourteen-in-One GeneX viral-based RNA-Targeted small Unique antiviral peptide Drug to treat severe cases of PAN-COVID.

III)Development of Alpha, Beta, Gamma, Delta, lamda & Omicron 3rd Fifteen-in-One GeneX viral-based RNA-Targeted small Unique antiviral peptide Drug to treat severe cases of PAN-COVID.

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Novelty of Research Dengue virus type 1-4

Since an epidemic pace-related rare disease, including dengue fever, are major threats to our society, In this project proposal aims at the development of generally applicable technologies bioinformatics, solid phase peptide synthesis (SPPS), biotechnology & molecular biology that will be applied to a novel target EDIII 1-4 protein, currently it is not in the market to facilitating novel anti-peptides monoclonal antibody for therapeutics. Synthesing of amino acids 7-44 length that result, peptide, that results in high purities with specific targets for the treatment of dengue fever. Since dengue fever is a major threat to our society, the novel peptides effective anti-peptide powder based form on specific targets based EDIII 1-4 produce different type of anti-peptide monoclonal antibody. It may alter the biosynthetic pathway, weather they produce new pharmacologically active compound natural products from readily available natural amino acids through solid phase or solution phase chemistry product based synhetic peptide powder form based anti-peptide antibody from EDIII 1-4 proteins. To develop new innovative interdisciplinary strategies to open new avenues for the development of a unique specific peptide sequence to be used for the rabbit immunization for monospecific antibody and synthetic peptide powder-based form anti peptide antibody against recombinant dengue virus antigen suitable for the treating of dengue fever is key to animal and future patients’ treatments and survival. Based on current investigations of the functional significance of dengue its potential role in dengue western blot and immunocytochemistry with fixed cell from a cell line but not for any in vivo experiment and also not for cell-injection in cell culture etc. we shall focus on developing synthetic peptide powder-based form anti peptide antibody against recombinant dengue virus antigen suitable for the treating of dengue fever in human. In this present proposal help to address the sustainable use of EDIII 1-4 proteins.
Current State-Of-The-Art Problem: DENGUE VIRUS 1-4

We put emphasize that with this computationally developed, predicts protein sequence while the invention of new novel study suggest that thymine content (uracil) in protein coding ssRNA sequences are randomly distributed. Thymine content distribution is used here to examine DENGUE (DENV 1-4). Frames are arriving as expected, positive single-stranded RNA (+ssRNA) first three frames, in the DENGUE VIRUS genome.

In this study suggest that protein coding DENGUE VIRUS 1-4 (DENV 1-4) the frequency on XTX, XAX, XCX and XGX of ORFs shown the DENV-1 and DENV-3 protein expressed on XTX frame 2 , DENV-2 protein expressed on XTX frame 1 and DENV 4 protein expressed on XTX frame 3 meanwhile XAX also shown the DENV1 and DENV3 protein expressed on XAX frame 2 , DENV2 protein expressed on XAX frame 1 and DENV 4 protein expressed on XAX frame 3 in low frequency and other codons of frames XGX and XCX not shown the ORFs results.

Need for the Study

Recent data show that the number of people affected by dengue is growing at an epidemic pace with an approximate annual increase nearly 390 million dengue infections yearly worldwide and up to 96 million dengue cases. To identification and development of a unique peptide sequence and synthetic peptide powder form based from EDIII 1-4 and Development of a unique synthetic peptide powder-based form to be used for production and characterization of monoclonal monospecific anti-peptide antibodies against recombinant dengue virus antigen suitable for the treating of dengue fever will be of highest significance as well.

Proposed Solution-DENV 1-4 by LYSINE BIOTECH

Our computational studies provide new insight into the various types of unique specific DENGUE 1-4 protein coding analyzed frequency data on XTX, XAX, XGX and XCX frames 1-3 results were showed the DENV-1 and DENV-3 protein expressed on XTX frame 2 and DENV-2 protein expressed on XTX frame 1 and DENV 4 protein expressed on XTX frame 3 to compared other programme example JColorGrid software for the visualization of biological measurements for bioactivity data results in read will shown exactly our results from the results our Protein coding frmaes programme will shown a very good results for bioactivity data of DENGUE 1-4

LYSINE BIOTECH will be granting to in-license CDSCO/FDA Indian rights of four in one EDIII type 1-4 GeneX viral-based Recombinant peptide vaccine candidates demonstrating potential efficacy against DENV 1-4
LYSINE BIOTECH will be granting to in-license CDSCO/FDA Indian rights of EDIII type 1-4 tetra-specific GeneX viral antipeptide viral Drug candidates with strong binding targeting DENV 1-4

Specific Goals of Research

Protein sequence target specific (N-terminal and C-terminal) based on antibody epitope prediction for rapid test antigen and antibody to peptide/protein drugs up to vaccines for DENV 1-4
Recombinant Protein Expression and Purification of four in one EDIII type 1-4 proteins sub-unit vaccine candidates for DENV 1-4 Platform/technology from Yeast, CHO Cells and Insect Cells
BIRAC-NBM: proposal no. BT/NBM0103/02/18 submitted by LYSINE BIOTECH, ICMR-NIE CHENNAI and TANUVAS-CHENNAI entitled "Development of monoclonal antibodies from envelope protein of Dengue virus, novel detection assay for the early diagnosis and future explorations on therapeutic applications". The proposal could not qualify the highly competitive Technical Evaluation Process of NBM.
Chemical synthesis and peptides drug and vaccines are usually composed of 7–44 amino acids containing the EDIII type 1-4 specific epitope of an antigen for DENV 1-4
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Publications

I).Mayakrishnan Vijayakumar, BalakarthikeyanJanani, PriyaKannappan, Senthil Renganathan*, SameerAl-Ghamdi, Mohammed Alsaidan, Mohamed AAbdelaziz, Abubucker Peer Mohideen, Mohammad Shahid, Thiyagarajan Ramesh, 2021.In silico identification of potential inhibitors against main protease of SARS-CoV-2 6LU7 from Andrographis panniculata via molecular docking, binding energy calculations and molecular dynamics simulation studies Saudi Journal of Biological Sciences. Oct.29-2021. doi: 10.1016/j.sjbs.2021.10.060.

II).Renganathan Senthil*, Manokaran Sakthivel, Singaravelu Usha, 2021. Structure-based drug design of peroxisome proliferator-activated receptor gamma inhibitors: ferulic acid and derivatives. J Biomol Struct Dyn. 39(4):1295-1311.doi: 10.1080/07391102.2020.1740790.

III).Konda Mani Saravanan, Haiping Zhang, Renganathan Senthil*, Kevin Kumar Vijayakumar, Vignesh Sounderrajan, Yanjie Wei, Harshavardhan Shakila, 2020. Structural basis for the inhibition of SARS-CoV2 main protease by Indian medicinal plant-derived antiviral compounds. J Biomol Struct Dyn. 1-9.doi: 10.1080/07391102.2020.1834457.

IV).Mahendrarajan Venkatramanan, Pitchaipillai Sankar Ganesh, Renganathan Senthil*, Jeyachandran Akshay, Arumugam Veera Ravi, Kulanthaivel Langeswaran, Jamuna Vadivelu, Samuthira Nagarajan, Kaliaperumal Rajendran, Esaki Muthu Shankar, 2020. Inhibition of Quorum Sensing and Biofilm Formation in Chromobacterium violaceum by Fruit Extracts of Passiflora edulis. ACS Omega.5(40):25605-25616.doi: 10.1021/acsomega.0c02483.

V).Subrata Pramanik, Manisha Thaker, Ananda Gopu Perumal*, Rajasekaran Ekambaram, Naresh Poondla, Markus Schmidt, Pok‐Son Kim, Arne Kutzner, Klaus Heese, 2020. Proteomic Atomics Reveals a Distinctive Uracil‐5‐Methyltransferase. Molecular Informatics. Vol. 39, Issue 5,doi:10.1002/minf.201900135 .

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