Products Development Programs in cGMP Facility
To secure and ensure long term financial support, VRDC has considered using the current vaccine product development programs to establish the cGMP Facility as a Technology Transfer unit. Since 2004 VRDC has utilized the funding from the Taiwanese government to establish the infra-structure of cGMP Facility from ground zero to now with a critical mass of >100 staff who are well trained to produce traditional vaccine like BCG and GMP-grade vaccine candidates for clinical trials
With well-trained GMP staff, the completion of the infra-structure and facility validation, VRDC cGMP pilot plants now play very important and leading role in Taiwan vaccine industry (see the diagram below). In the past, the lack of GMP pilot plants and experienced production staff, it has been difficult to evaluate any potential vaccine candidates developed by Taiwan Universities and/or local Biotech for their efficacy in human phase 1 clinical trials. In fact the Vaccine Center had demonstrated her values. During the H1N1 pandemic crisis in 2009 the Vaccine Center had provided contract-services to assist a local Biotech Company to manufacturing emergency H1N1 flu vaccine candidates for phase 1 human clinical trials.
In 2007 we have initiated a “Human Vaccine R&D programs funded by DoH/NSC to develop 3 vaccine candidates and complete their phase 1 clinical trials by the end of 2011. We submitted Investigational New Drug (IND) dossiers to Department of Health CDE-98IND06088) for initiating phase 1 trail of cell-based H5N1 flu vaccine candidate (ProVEU) to be conducted in National Taiwan University Hospital (NTUH) in July/2009 that was approved by Taiwan FDA (#0980324995) in Nov/2009. This was the first Taiwanese developed cell-based H5N1 flu vaccine candidate tested in phase 1 clinical trial. The trial was completed in July 2010 and this vaccine candidate was proven to be safe and could elicit anti-H5N1 flu antibody responses. To obtain the global exclusive product licensures and marketing rights, a local Biotech company Medigen had paid the up-front money and agreed to milestone payment worth >30 millions NTD. Similarly, EV71 vaccine candidate (EV71vac) was approved by Taiwan FDA (#0991413522) for phase 1 trials in Oct 26/2010, and completed the first stage of 10 volunteers’ immunization in June/2011. We did not observe any side-effect, but obtained excellent virus neutralization titers from these volunteers. We are currently signing EV71 vaccine non-exclusive licensing agreements with two local companies. The 3rd vaccine candidate recombinant lipoprotein of meningococcal group B (MGBvac) has just completed the pre-clinical toxicology study. The IND has been filed to TFDA and most likely will enter phase 1 trial in Q3/2011. All these potential vaccines could be financially attractive for a spin-off business unit in the Vaccine Center.
Specified Program: Taiwan Human Vaccine R&D Program
In order to implement the policy of the government for developing vaccine self-manufacturing capability and strengthening the infra-structure of vaccine research, and also to responding to any threat from the emerging infectious diseases, this project will integrate all the resources and expertise in all fields to establish an national vaccine research and development team dedicating for the local important vaccine research and production.
The epidemic of influenza is now a national public health concern, and the World Health Organization is encouraging every country to strengthen the pandemic influenza preparedness and responses, in particular on vaccine research and development and to build up platforms that have the capability of self-manufacturing. The enterovirus type 71 can cause severe neurological disease, and even death. The mortality rate in Taiwan is particularly high compare to other countries, so it is one of the targeted vaccines focused by Vaccine Center. The drug resistance of Meningococcus group B is also causing a threat to the global health, and from our preliminary animal protection studies data; it has shown that the subunit vaccine for Meningococcus group B is feasible.
Based on public health concerns, disease prevention and to have the ability to response to any pandemic outbreak, H5N1 influenza, enterovirus types 71, and meningococcal recombinant vaccine are the targeted vaccines to be developed in this program. The ultimate goal of this project is to prepare and to enter the phase I clinical trial for 3 of these targeted vaccines in five years time (Jan/2007-Dec/2011).
1. MDCK Cell-based H5N1 influenza vaccine (ProVEU) project：
In 2010, we had
- completed the immunization of H5N1 candidate vaccine, ProVEU with 60 test subjects for phase I clinical trial, found the ProVEU to be safe and immunogenic in human.
- performed two test runs of 50 liter bioreactor (30 L working volume) microcarrier H5N1 flu virus production, the HA titer were found to be 512~1024/50μL and base on this titer, 100,000 doses of 15 mcg HA per year; one run of 150 liter bioreactor, (120 L working volume) the HA titer was determined to be 512/50μL and base on this titer, 150,000 doses of 15 mcg HA per 3 months;
- Signed cell-based H5N1/H1N1 flu vaccine technology transfer with a local company with up-front and milestone payments worth >30 millions NTD;
- a novel adjuvant discovery for flu vaccine, found that the presence of D-form stereoisomer in the native-peptide sequence of indolicidin could increase the resistance of peptide segment to hydrolyze. The secretion of IFN-γ (a predominant T helper type 1 cytokine relevant to virus-specific CTL activity) in the LD-indolicidin group was significantly higher than those in the non-adjuvanted or indolicidin-adjuvanted group. Therefore, antigen formulated with LD-indolicidin could enhance the T-cell responses
- found that the mice received PELC/CpG formulated-vaccines could generate effective cross-clade neutralizing antibodies against H5N1 flu virus. Thus, PELC/CpG combination adjuvant is an effective tool in pre-pandemic preparedness and antigen sparing.
- found that the mice received H5N1 inactivated virus vaccine could not generate cross-neutralizing antibodies against H1N1, even in the presence of alum adjuvant. Similarly, the mice received H1N1 virus could not generate cross-neutralizing antibodies against H5N1. The use of combo vaccines is a practical way to generate neutralizing antibodies against H1N1 and H5N1 simultaneously. One should keep in mind that a combo vaccine is not a procedure just to put the antigens together. New adjuvants will be requested to overcome antigenic competition since there are limited receptor numbers of the immune cells. Our data showed that PELC has potential of overcoming antigenic competition and broaden the neutralizing antibody responses, in comparison with alum adjuvant. On the other hand, antigen formulated with LD-indolicidin only could enhance the T-cell responses but not the humoral protective antibodies, in comparison with non-adjuvanted vaccine (p<0.05).
In 2011, we have:
- established the serum-free bioprocess using 30/150 liter microcarrier bioreactors for H5N1 influenza virus production in the cGMP facility; 150,000 doses of 15 μg HA estimated could be prepared per 3 months base on the current HA titer；
- collaborated with a Taiwan local company, Medigen Biotechnology Corp. to scale up the H5N1 influenza vaccine production using A Tide-cell disposable bioreactor system. According to the service-contract agreement, VRDC offered the cGMP facility and the technical service of downstream purification；
- 3 peer-reviewed papers are published, and 2 new patent applications were file.
- One patent that was filed in 2008, was granted by US PTC office.
2. Vero cell-based Enterovirus 71 vaccine (EV71vac) project：
In 2010, we had:
- completed 4 lots of vaccine bulk and 2 lots of vaccine product filled with clinical trial materials；
- established all SOPs of QC tests and their qualifications；
- received the validation reports of master cell bank、working cell bank and end of production cell. The reports showed that there was no contamination found in the cell bank. The oncogenicity study was completed in April and no oncogenic properties were found in end of production cells；
- completed the validation of both master and working virus seeds. The reports provided that there were no contamination was found in the virus seeds；
- completed the pre-clinical animal toxicology studies in both rat and rabbit models. It showed that there was no adverse event cause by EV 71 vaccine；
- submitted the CMC dossier and obtained the permission issued by TFDA. The phase I clinical trail was initiated at Taipei Veterans General Hospital in Dec., 2010.
In 2011, we have:
- performed 1 test run of 50 liter serum-free microcarrier bioreactor. The total purified viral protein was determined to be 62.57 μg/ml and base on this, 5,800 dose of 10 ug total protein per run；
- assisted Technology Transfer Office of NHRI who is currently negotiating with Medigen Biotechnology Corp and Adimmune Corporation about non-exclusive licensing agreement;
- completed the first stage of phase I clinical trial with 10 subjects at Taipei Veterans General Hospital in June;
- 5 papers published in peer-reviewed journal and 2 new patent applications filed.
3. Meningococcal group B recombinant vaccine (MGBvac) project：
In 2010, we had:
- filed a patent application covering the purification method of recombinant lipoprotein (USPTO Application No. 61/372,617)
- received the National Innovation Award for this recombinant lipoprotein technology at the meeting of the Institute for Biotechnology and Medicine Industry on May 17, 2010.
- The U.S. patent of “Lipidating Sequences and Use Thereof for Producing Lipidated Proteins in E. coli” was granted on November 16, 2010
- established a cGMP-certified facility for manufacturing the prototype vaccine of MGBvac
- a kanamycin resistant E. coli expression system developed to produce the recombinant Ag473 (antigen component of MGBvac) in a 20L fermentation scale. At least 7 batches of upstream processes and 3 batches of downstream processes have been conducted to demonstrate the robustness and the quality of production.
- The validations of cell banks including MCB, WCB, and end of production cells were conducted by the Bioreliance UK, according to the international regulations and guidelines；
- The identity, purity, and stability of MGBvac were examined by the state of the art technologies, including the GC- and LC- mass spectroscopes, etc. All the QC tests for the product release have been qualified and implemented
- For supporting the preclinical toxicology study and clinical phase I trials, one batch of MGBvac (Lot No. 40-06-0101) was prepared and filled as a multiple-dose formulation (3 mL/vial, 20 mcg/0.5 mL per dose)；
- The experimental design of animal pre-clinical toxicology studies in rat and rabbit models was reviewed by CDE (Center for drug evaluation, Taiwan) and performed by DCB
In 2011, we have:
- Documentations of Investigation New Drug (IND), pre-clinical studies, QC biochemical characterizations of vaccine, and animal toxicology studies of MGBvac performed, completed and submitted to CDE/TFDA for the rolling review
- The IND( Part I: Drug Substance and Part II: Drug product) file has been delivered to the CDE for review and scheduled to be completed before the end of August
- The clinical trials are expected to start after the IND submission and approval by Department of Health (DoH) in Q4/2011
- the bidding process for announcement for selection ” MGBvac” technology transfer listed in NHRI website July22/2011
- several articles published in the peer-reviewed journals and new patent application filed
4. Responses to Taiwanese Government Emergency Request of Vaccines
Cell-based Pandemic Flu Vaccine Development
H5N1 Pandemic Flu Vaccine Development (2007-2012)
- Completed MDCK cell banks and master virus seed bank of H5N1 vaccine strain RG14 validations
- Completed 3 cGMP clinical production batches with serum-free media
- Completed Pre-clinical toxicology studies, general safety test and vaccine stability program
- Phase 1 clinical trial IND submitted to DoH and approved in Aug/2009
- Phase 1 clinical trial is completed in Q2/2010 at NTUH, showed to be safe and immunogenic
H1N1 Pandemic Flu Vaccine Development (June/2009 – 2011)
- Completed the production of 3 batches of 60 liters using roller bottle technology
- Cell-based H1N1 Vaccine candidate is tested and ready to be used
- Pre-clinical general safety test and stability have been completed
- 30 liter and 150 liters Micro-carrier bioreactors are established for virus production
- Currently assisting Medigen to produce H1N1/H5N1 vaccine candidate using a signed service contract
- Exclusive Technology Transfer of MDCK cell-based flu vaccine technology is signed with local Medigen Biotech