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Cancer Drug Slows Poxvirus in Mice
http://www2.niaid.nih.gov/Newsroom/Releases/cancer_pox.htm
Mice given a relatively new cancer drug can survive an otherwise lethal dose of vaccinia virus, a relative of smallpox virus, report scientists supported by the National Institute of Allergy and Infectious Diseases (NIAID), part of the National Institutes of Health. The findings, say the investigators, suggest that Gleevec or similar drugs might be useful in preventing adverse side effects of smallpox vaccine. The classic smallpox vaccine is made from live, weakened vaccinia virus and is not recommended for people with compromised immunity, except in emergency situations where they may have been exposed to smallpox virus.
“This study helps illuminate the cellular machinery used by poxviruses to exit infected cells, and also provides new support for the concept of treating viral infections by targeting specific host cell molecules rather than the viruses themselves,” says NIAID Director Anthony S. Fauci, M.D.
The senior author of the paper, published online this week in the journal Nature Medicine, is Daniel Kalman, Ph.D., of Emory University School of Medicine in Atlanta.
Like all viruses, poxviruses co-opt various cellular molecules and processes to enter a cell, replicate and then spread to uninfected cells. Using lab-grown cells, Dr. Kalman and his colleagues identified specific cell proteins vaccinia uses to detach from an infected cell and move toward an uninfected cell. The proteins, members of the Abl-family (pronounced “able”) of tyrosine kinases, are well known to cancer investigators because mutation of one family member, Abl, causes a rare form of cancer known as chronic myelogenous leukemia (CML). Gleevec inhibits Abl-family tyrosine kinases and has proved very useful in treating CML.
To learn whether Gleevec could prevent or lessen vaccinia’s ability to spread in a living organism, the researchers treated mice with either saline solution or with Gleevec at a dose equivalent to that given to humans being treated for CML. Next, they exposed the mice to ordinarily lethal amounts of vaccinia. All of the Gleevec-treated mice survived, while 70 percent of the untreated mice died.
This finding, if confirmed in additional animal model studies, suggests that Gleevec might play a role in addressing a public health emergency in the event of a smallpox outbreak, Dr. Kalman says. Specifically, Gleevec might be useful as a preventative against adverse effects of smallpox vaccine, enabling clinicians to use the vaccine even in people who otherwise could not take it. Given for a short period, Gleevec theoretically could hamper the cell-to-cell spread of virus and allow the body’s immune system to mount a successful defense, he explains. Experiments to test whether Gleevec might work against smallpox virus as well as against vaccinia virus are now being planned, Dr. Kalman says. “The approach of fighting disease by targeting drugs to cellular molecules rather than to disease agents themselves may be applicable to a wide variety of pathogenic microorganisms,” he says.
Routine vaccinations for smallpox ended in this country in the early 1970s, and the World Health Organization declared smallpox eradicated in 1980. Nevertheless, concern remains that smallpox virus could be unleashed through an act of bioterror. For this reason, scientists are working to better understand the mechanisms of smallpox disease and to develop new and improved smallpox treatments and vaccines.
NIAID is a component of the National Institutes of Health, an agency of the U.S. Department of Health and Human Services. NIAID supports basic and applied research to prevent, diagnose and treat infectious diseases such as HIV/AIDS and other sexually transmitted infections, influenza, tuberculosis, malaria and illness from potential agents of bioterrorism. NIAID also supports research on transplantation and immune-related illnesses, including autoimmune disorders, asthma and allergies.
###
Reference: PM Reeves et al. Disabling poxvirus pathogenesis by inhibition of Abl-family tyrosine kinases. Nature Medicine DOI: 10.1038/nm1265 (2005).
GlaxoSmithKline to Launch 5 Major Vaccines
Thursday June 30, 8:47 am ET
GlaxoSmithKline Expects to Launch Five Major Vaccines in the Next Five Years
NEW YORK (AP) -- British drug maker GlaxoSmithKline PLC said Thursday that it has more than 20 vaccines in clinical development, with five major launches planned in the next five years.
The company said it will launch Cervarix for the prevention of HPV which causes cervical cancer, Rotarix for rotavirus gastroenteritis, Streptorix for pneumococcal disease, an improved flu vaccine and Hib-MenCY vaccine combinations against meningitis.
These five products will enter markets that have the potential to reach a total market value of $11 billion to $18 billion by 2010, Glaxo said.
The company estimates that cumulatively, up to 80 million women could be vaccinated by 2010 for protection against cervical cancer, and the cervical cancer vaccine market could be valued at $4 billion to $7 billion per year by 2010. Regulatory filings of Cervarix will take place in Europe in the first half of 2006, and in international markets during 2006. Discussions on filing with the Food and Drug Administration are ongoing in the U.S., where the vaccine has been granted "fast track" status.
Glaxo said the launch of its new pediatric vaccines, combined with its existing portfolio, will give the company a full range of products to meet all infant vaccination schedules in all major markets within the first six months of life.
Glaxo estimates that the global market for rotavirus vaccines could be valued at $1.8 billion to $2.4 billion by 2010. GSK has submitted regulatory filings for Rotarix in Europe and 50 other markets, and Rotarix has now been approved in 7 markets, including Mexico. Discussions on the U.S. filing are ongoing with the FDA.
Glaxo also said it plans to double its Fluarix manufacturing capacity to 80 million doses annually by 2008, is developing an improved flu vaccine using a unique adjuvant. The drug maker will develop a new cell culture manufacturing process by 2010 to further boost production capacity.
Glaxo estimates that the influenza market could more than double from its current value to $2.9 billion to $3.7 billion in 2010. Fluarix is licensed in 102 countries with 2004 sales of $144 million, and the company filed Fluarix with the FDA in May in time to be available for the 2005 to 2006 flu season, if it is approved.
Large Phase III trials of the improved flu vaccine are due to start in 2006, and regulatory filings will take place in the U.S., Europe and international markets in 2008, the company said.
GlaxoSmithKline Biologicals, the company's vaccine manufacturing unit, is located in Rixensart, Belgium and posted 2004 sales of nearly $2 billion. In 2004, GSK Biologicals distributed more than 1.5 billion doses of vaccines to 168 countries in both the developed and developing world. About 140 million of the doses delivered last year were combined pediatric vaccines that protect children against a minimum of three and up to six diseases in one vaccine.
jake, big difference betweeen
vaccine, and therapeutic!
j
ms kso- thanks, interesting. Another testament to the very safe profile of tarvacin compared to what little competition is out there in the anti-viral realm.
on another topic-
I'm trying to find info on the "extension protocol" in the cancer trial... My hope is that it will be more than additional available montherapy beyond the ten week course of the trial.
We're both very familiar with the statistics on mono & combo.
Mono was quite impressive, but combo with chemo or radiation was spectacular. As we all know folks who may qualify for the trial, I'd like to get this answer so I can better inform people.
I've spoken with Ken at Hawk, and emailed him my question, hoping to hear back soon. If you have any info in this area I'd appreciate you emailing it etc.
j
Animal Rule
http://www.fda.gov/cber/summaries/biochem042005ck.pdf
• To reduce or prevent serious or life threatening
conditions caused by exposure to lethal or
permanently disabling toxic chemical, biological,
radiological, or nuclear substances.
• Expected to provide meaningful benefit over
existing therapies.
• Human efficacy trials not feasible or ethical.
• Use of animal efficacy data scientifically
appropriate.
• Does not apply if approval can be based on efficacy
standards elsewhere in FDA regulations.
Animal Rule (cont.)
• Still need human clinical data:
– PK/immunogenicity data, and
– Safety in population(s) representative of use.
• Civilian use often includes pregnancy, children.
• Approval subject to post-marketing studies and/or
restrictions on use.
• Please work closely with FDA on planning animal
studies before starting them.
• Potential limitations:
• Where there is no valid animal model of disease;
• How to predictably bridge animal data to humans; and
• Confidence may be an issue, even in valid models.
• Examples of current studies on threat pathogens:
– Smallpox - assays for immune response and potency,
vaccine safety (neurovirulence), risk assessment on
vaccine strategies and blood safety....
----------------------------------------------
Steve King:
"....tarvacin has shown activity in the treatment of animals infected with lethal doses of lassa fever which is a category-A bio-terrorism watchlist virus."
...."Our plans for developing the Tarvacin antiviral program are really going in two different directions. The first is really geared toward bio-defense applications, so this could be either bioterrorism applications or widespread viral outbreaks. Our priority is to generate preclinical data. As the previous speaker mentioned there's really no way to run clinical trials for many of these bioterrorism threats, so collecting preclinical data is really about the best you can do. We then want to combine that with human safety studies that we will be generating in the Hepatitis C clinical trial, as well as our anti-cancer clinical trial, and we believe- pending positive results of course- that this could be a fairly rapid route to regulatory approval for tarvacin for the treatment of- again- widespread viral outbreaks."
...."In April of this year, Peregrine and NIAID entered into an Agreement to screen tarvacin for activity against a broad spectrum of enveloped viruses, and these really fall into two categories- those of health concern, and those of bio-terrorism concern."...
"So to kind of summarize why we're excited about tarvacin in particular for the bio-defense applications:
Number one- we've received promising preclinincal results in the lassa fevr animal model. Those included both short-term treatment advantages, as well as long term immunity to the viral infection.
The second is we have current collaborations with NIAID that are underway to screen against viral bio-terrorism and health threats. This is quite an extensive list of viruses they're testing the drug against. In fact, their indication to us was that this is one of the first times they've ever tested such a broad spectrum of viruses with a single agent.
Thirdly is discussions are taking place to screen tarvacin for activity against additional bio-terrorism and health threats again these would fall under the marburg and ebola virus families.
Fourth, data from preclinical human phase one safety could be used to support expedited FDA approval as a therapy for bio-terrorism and health threat concerns, and this kind of falls under the animal rule which was recently enacted and implemented by the FDA, in which if you can combine good preclinical efficacy data along with good human phase one safety data, then you can effectively go directly for the approval process.
Fifthly, the mechanism of action and target indicates potential as a single agent that can be effective against a wide number of enveloped viruses, including those that are known about and those that are not known about,
and, lastly- human clinincal studies are not far away. They will be beginning over the short term. This will provide us with critical- number one- safety data for the program, which will be necessary for any future approval filings, but also, in those same clinical trials we'll be looking for the effectiveness of removing the hepatitis C virus by looking at viral load changes."....
Steve King, president/CEO
--------------------------------------
National Institute of Allergies and Infectious Diseases to Test Tarvacin(TM) for Antiviral Potential
Monday April 4, 8:30 am ET
- Peregrine Pharmaceutical's Anti-Phospholipid Therapy to Be Screened Against Broad Spectrum of Viruses That Threaten Worldwide Health and Global Security
TUSTIN, Calif., April 4 /PRNewswire-FirstCall/ -- Peregrine Pharmaceuticals Inc. (Nasdaq: PPHM - News) and the National Institute of Allergy and Infectious Disease (NIAID) have agreed to a collaboration in which NIAID's testing laboratories will screen Peregrine's Anti-Phospholipid Therapy agents, including Tarvacin(TM), for activity against a broad spectrum of enveloped viral pathogens of health and bioterrorism concern. Virus types to be screened as part of the collaboration potentially include herpes viruses, respiratory viruses, pox viruses, Hepatitis B and C, Papillomavirus and viruses of biodefense concern including Pichinde, Yellow Fever, West Nile and Dengue.
"We are very pleased to be able to work with the NIAID to further explore the anti-viral potential of Tarvacin(TM) and the rest of our Anti-Phospholipid Therapy agents," said Steven King, president and CEO of Peregrine Pharmaceuticals. "The data that will be generated from this collaboration will be very helpful in guiding development of our Tarvacin(TM) anti-viral clinical program."
During the first year of an ongoing 3-year, $1.68 million grant from the NIAID, scientists at the University of Texas Southwestern Medical Center at Dallas determined that Tarvacin(TM) binds to virally infected cells and viral particles including Pichinde virus, which causes a fatal viral hemorrhagic fever and is used as an established model for Lassa fever. Pichinde virus is on the U.S. government's biodefense Category A watch list. The researchers further determined that Tarvacin(TM) also significantly protected animals challenged with a lethal dose of Pichinde virus. Results from these studies are being presented at the annual meeting of the American Association of Immunologists taking place this week in San Diego, California.
Pathogens to be screened in this collaboration belong to a class of viruses known as "enveloped viruses," which derive their outermost coating from their host cell membrane during viral replication. Enveloped viruses account for many of the most concerning viral health risks including HIV, Hepatitis B and C, cytomegalovirus, hemorrhagic fever, SARS and various types of influenza including Avian influenza.
About Anti-Phospholipid Therapy in the Treatment of Viral Diseases
Anti-Phospholipid Therapy is Peregrine Pharmaceuticals' novel approach to treating cancer, viral infections and certain ocular diseases. It is based on the finding that aminophospholipids, which are basic components of the inner surface of the cell membrane, become exposed as antigenic targets in certain disease states.
A large number of viruses that impact global health and security possess an "envelope" derived from their host cell membrane. Since viruses lack the means to maintain structural organization of the envelope, amino-phospholipids such as phosphatidylserine (PS) and phosphatidylethanolamine (PE) become exposed on the surface of these viruses, making them potential therapeutic targets. Peregrine Pharmaceuticals, together with its collaborators, has developed a series of monoclonal antibodies directed against aminophospholipids to take advantage of this property.
this quote from Thorpe I copied from cjgaddy's intro above may hint at a bit of what me may learn about tomorrow- cytomegalovirus.
Thorpe: (re: Lassa experiment)....."That's an extraordinary result, I don't know of any other antiviral agent that is known to protect against Lassa Fever. And we've also shown similar results in cytomegalovirus... at that dose we've never seen any sign of toxicity in mice or monkeys; about 14 species treated with the therapeutic dose. And that's thousands of mice & monkeys. And even if you increase the dose to 10 mg/kg, that's 10x the therapeutic dose.. So the conclusions with Tarvacin are that it has a unique mechanism of action, there's nothing else like it out there"....
EZ, yeah, PPHM on reg SHO list today-
http://www.nasdaqtrader.com/aspx/regsho.aspx
I thought they already 'dealt away' VEGF trap a couple years ago to Aventis for up to ~ 1/2 billion. ?
j
ms kso, regarding the VX-950 Phase 1b trial: it's the amount of the doses, their frequency, and the rise of the virus count after stopping treatment, that makes me not too enthused about VX-950. And- at the conclusion of 1b they mention,
"combination therapy wih other antiviral agents or IFN may be required to obtain optimal responses"
My mother did a year or so of interferon (for melanoma), and she did not have much fun... Nasty stuff. I hope for the sake of those suffering with HCV that the VX-950 monotherapy works well enough on it's own.
The phase 2 VX-950 trial should shed more light on this angle.
And any HCV info from Peregrine on Wed. will be very interesting to compare (if at all possible) with the early VX-950 data.
j
ms kso, agreed, BOTH the Peregrine P1 trials are saving a year or so, I assume due to really really nice looking preclinical data.
j
ms kso, more vertex-
VX-950
phase 1a- (healthy subjects only for PK data etc.)
October 9, 2004
Phase 1a Study of Oral HCV Protease Inhibitor Complete
Vertex Pharmaceuticals recently completed the dosing portion of a phase 1a clinical trial on an investigational oral protease inhibitor for the treatment of the hepatitis C virus (HCV) infection. The study, conducted in Europe on healthy volunteers, assessed safety, tolerability and pharmacokinetics in escalating, single doses of the drug, called VX-950. Doses ranging from 25mg to 125mg were administered. No dose-limit toxicities were identified and a maximum tolerated dose was not reached, but blood concentrations were observed that exceeded the concentration known to demonstrate potential antiviral activity in preclinical laboratory experiments. Based on these results, Vertex expects to begin a phase 1b clinical study in HCV-infected patients by the end of the year.
phase 1b Nov 11, 2004
Vertex begins phase Ib hepatitis C trial
Global biotech firm Vertex Pharmaceuticals (NASDAQ: VRTX - news) , a small-molecule drugs developer, has initiated a phase Ib clinical trial for its investigational oral protease inhibitor for the treatment of hepatitis C infection.
Global biotech firm Vertex Pharmaceuticals, a small-molecule drugs developer, has initiated a phase Ib clinical trial for its investigational oral protease inhibitor for the treatment of hepatitis C infection.
The double-blind, placebo-controlled study will evaluate the tolerability, pharmacokinetics and viral kinetics of multiple, ascending doses of VX-950 over a period of up to 14 days and will enroll approximately 60 subjects. This is the first reported initiation of a study that involves 14 days of administration of a hepatitis C virus (HCV) protease inhibitor in patients with chronic hepatitis C infection.
The clinical study will first assess healthy volunteers receiving multiple doses of VX-950 for a five-day period. Following this initial assessment, the study will evaluate three different doses of VX-950 in HCV patients, over 14 days of treatment in serially configured dose groups.
Pre-clinical studies have shown that VX-950 significantly reduces levels of HCV RNA in both an in vitro replicon system and infectious virus assays. In the phase Ia clinical study, VX-950 was well-tolerated and demonstrated oral bioavailability.
Furthermore, combined clinical and pre-clinical pharmacokinetic results indicate that VX-950 can be administered in a dose regimen that may achieve liver concentrations substantially greater than target concentrations (IC50 and IC90 in non-clinical studies).
"The phase Ib study is expected to provide us with important information on the ability of VX-950 to reduce viral load, which we expect will help to define the potential clinical impact of this new class of drugs," said Dr John Alam, senior vice president of drug evaluation and approval at Vertex.
Vertex anticipates that the phase Ib study will be completed in the first half of 2005.
phase 1b complete:
Number of Subjects
Headache: 5 (28%)
Diarrhea:3 (17%)
Nausea: 2 (11%)
frequent urination: 2 (11%)
sleepiness/drowsiness: 2 (11%)
rapdity of viral load decline, undetectable HCV RNA & slow return of HCV RNA post-dosing suggest that VX950 should be explored as a monotherapy
--combination therapy wih other antiviral agents or IFN may be required to obtain optimal responses
--treatment duration with HCV PI therapy may be shorter than the current standard of care
I also look forward someday to seeing the first mention of in vitro or mouse anti-bacterial infection data, or parasitic infection, (as mentioned in Thorpe's APT patent applications).
One of these days. "lantibiotics"...
j
kso,
Yup. HCV data for sure IMO. I also hope/expect to see some HIV and flu data this week.
King in nyc: ..."In addition we are developing tarvacin for common viral applications. There, we've collected preclinical data on hepatitis C virus, we're continuing studies with HIV, influenza, and we're looking at some other potentials there. We will then complete human clinical studies for each of those viral types".
I assume we'll be seeing some darn nice Hepatitis C data.
and to think- the trial should be getting underway soon. It's cheap- same drug as other trial, made in house, viral trial, most likely out-patients, shoot 'em up, take some blood every few days or once a week for a month or two...
IF it works, it has some pretty big implications-
* as a proof of principle for the mechanism of action implying
likely efficacy in ANY AND ALL enveloped viruses,
* as safety data to be used for NDA's(!) for a plethora
of bio-terror related viral treatments like ebola, marburg, hantavirus, smallpox, etc...
* and as groundwork for trials for a whole slew of
non-bio-terror related viral IND's like HIV,
influenza, Herpes, CMV, HPV, etc....
The thing is- once patients are dosed in this approved
HCV trial, the company will have info related to human
safety and efficacy in a matter of weeks/months.
And- as for the chances of it not working in humans as
it has worked in every other mammal, very safely and
effectively, due to the nature of the mechanism of
action,I have good reason to be optimistic about
similar efficacy-
Thorpe: The phospholipids that they recognize have the same
structure and cellular distribution in different
mammalian species, simplifying the transition from
experimental animals into humans. The antibodies are
not toxic to mice, even when administered in high
doses for prolonged periods of time.”
Steve King quotes from bio-defense speech:
(These are what stood out to me. I have left out info on various supporting material related to disease pandemics, and a lot of info regarding the tarvacin mechanism of action, as I think most are familiar with how it works and how broad it's potential is for treating both all cancers and all enveloped viruses, (not to mention non-enveloped viruses, bacterial infections, and also parasitic infections, according to other Thorpe patent applications, but that is a topic for another day )....
I highly recommend all who are interested to have a listen to King's talk. It's only 18 minutes, and it's right here:
http://www.wsw.com/webcast/rrshq6/pphm/
-------------------------------------------------------------------------------------
Steve King:
...."We believe we have an agent that has the potential to have some broad utility in the bio-defense area, whether it's bio-terorism or in the control of widespread viral outbreaks."
...."Tarvacin recognizes a stable target common to all enveloped viruses that cannot easily be mutated because it is derrived from the host, it's not encoded by the genome, but rather it's a stable structure of the virus particle. Examples of enveloped viruses include marburg virus, lassa fever, ebola, HIV, Hepatitis C, influenza. Secondly as I'll show you later in some preclinical data, tarvacin has shown the ability to arrest the development of disease even in late stages of infection. And thirdly- tarvacin is expected to recognize not just the known enveloped viruses that are here today, but new enveloped viruses that will undoubtedly become available and will have mutated and become new viral strains over the next years to come."
"This is an example of the enveloped viruses that are on the class-A watch list from the Center for Disease Control. You can see the viruses that are on here such as lassa fever, yellow fever, west nile virus, marburg virus...., - and the reason I put this up here is that we fully expect that tarvacin would be able to recognize all of these viral types, not just individual viruses where we'd have to develop new agents for each virus, but rather a single agent that could recognize all of these.".....
...."As a summary of the preclinical data that we've generated to date, we've found that tarvacin recognizes multiple enveloped virus strains. In fact- every enveloped virus strain we've tested to date. This data will be presented next week at Bio 2005."
Secondly, tarvacin recognizes virally infected cells from multiple virus types. This again has been shown in a number of systems and we're continuing those studies, and so far in every system we've looked at we've seen the expression of the tarvacin target on these virally infected cells.
Thirdly, tarvacin has shown activity in the treatment of animals infected with lethal doses of lassa fever which is a category-A bio-terrorism watchlist virus.
We've also seen even more significant anti-viral effects in other animal models, and again that data will be presented at the bio conference next week."
...."Our plans for developing the Tarvacin antiviral program are really going in two different directions. The first is really geared toward bio-defense applications, so this could be either bioterrorism applications or widespread viral outbreaks. Our priority is to generate preclinical data. As the previous speaker mentioned there's really no way to run clinical trials for many of these bioterrorism threats, so collecting preclinical data is really about the best you can do. We then want to combine that with human safety studies that we will be generating in the Hepatitis C clinical trial, as well as our anti-cancer clinical trial, and we believe- pending positive results of course- that this could be a fairly rapid route to regulatory approval for tarvacin for the treatment of- again- widespread viral outbreaks.
In addition we are developing tarvacin for common viral applications. There, we've collected preclinical data on hepatitis C virus, we're continuing studies with HIV, influenza, and we're looking at some other potentials there. We will then complete human clinical studies for each of those viral types, again- our hepatitis C clinical trial should be beginning soon, and we'll initiate new clinical trials as appropriate in timelines and of course this is what I would call a more standard route to regulatory approval"
...."In April of this year, Peregrine and NIAID entered into an Agreement to screen tarvacin for activity against a broad spectrum of enveloped viruses, and these really fall into two categories- those of health concern, and those of bio-terrorism concern."...
"So to kind of summarize why we're excited about tarvacin in particular for the bio-defense applications:
Number one- we've received promising preclinincal results in the lassa fevr animal model. Those included both short-term treatment advantages, as well as long term immunity to the viral infection.
The second is we have current collaborations with NIAID that are underway to screen against viral bio-terrorism and health threats. This is quite an extensive list of viruses they're testing the drug against. In fact, their indication to us was that this is one of the first times they've ever tested such a broad spectrum of viruses with a single agent.
Thirdly is discussions are taking place to screen tarvacin for activity against additional bio-terrorism and health threats again these would fall under the marburg and ebola virus families.
Fourth, data from preclinical human phase one safety could be used to support expedited FDA approval as a therapy for bio-terrorism and health threat concerns, and this kind of falls under the animal rule which was recently enacted and implemented by the FDA, in which if you can combine good preclinical efficacy data along with good human phase one safety data, then you can effectively go directly for the approval process.
Fifthly, the mechanism of action and target indicates potential as a single agent that can be effective against a wide number of enveloped viruses, including those that are known about and those that are not known about,
and, lastly- human clinincal studies are not far away. They will be beginning over the short term. This will provide us with critical- number one- safety data for the program, which will be necessary for any future approval filings, but also, in those same clinical trials we'll be looking for the effectiveness of removing the hepatitis C virus by looking at viral load changes."....
Steve King, president/CEO
katie,
re: "shown the ability to arrest development of disease even at late stages of infection"
I agree, a very important statement.
My thoughts on that-
In the only animal viral efficacy data I'm familiar with, Thorpe, PL, & SK have all mentioned repeatedly how sick the guinea pigs were at the time of infusion. They let them go very far with the infection- they had lost weight, they had super high fevers, their fur was visibly affected etc.. It may be that they were on the brink of absolutely nothing being able to save them. Tarvacin saved half of them, who went on to be immune to further infection, and to lead normal lives, (until I assume they killed them)...
It may be (I'm figuring) that in further experiments they've generated much better statistics on tarvacin than this first study, which in itself was unprecedented and obviously very impressive. I think we'll be seeing numbers closer to 100% in animal studies with the animals doesed with tarvacin less closer to death, dosed soon after infection with fewer or no physical signs of infection, OR possibly- studies to demonstrate immunity by dosing before infection, and further tests into how long immunity to any enveloped virus lasts after "inoculation"... (interesting hmm?)
I'm sure the govt. is investigating many angles, including not only therapeutic treatment after infection but also as a possible vaccine, and if so, due to the mechanism of action, this admitedly increasingly hypothetical vaccine would be for all enveloped viruses.
As I said, I look forward to next weeks data!
SK says, ""AS A SUMMARY OF THE PRECLINICAL DATA THAT WE'VE GENERATED TO DATE, WE'VE FOUND THAT TARVACIN RECOGNIZES MULTIPLE ENVELOPED VIRUS STRAINS, IN FACT - EVERY ENVELOPED VIRUS STRAIN WE'VE TESTED TO DATE. THIS DATA WILL BE PRESENTED NEXT WEEK AT BIO 2005. "
ALSO SHOWN TO RECOGNIZE INFECTED CELLS FROM EVERY ENVELOPED VIRUS STRAIN.
"WE'VE ALSO SEEN EVEN MORE SIGNIFICANT ANTI-VIRAL EFFECTS IN OTHER ANIMAL MODELS, AND AGAIN THAT DATA WILL BE PRESENTED AT THE BIO CONFERENCE NEXT WEEK."
"We have current collaborations with NIAID that are underway to screen against viral bio-terrorism and health threats. This is quite an extensive list of viruses they're testing the drug against. In fact, their indication to us was that this is one of the first times they've ever tested such a broad spectrum of viruses with a single agent."
IMO, the innoculation pre-infection angle is a biggie, even if the immunity only lasts a relatively short time, as it will have huge implications for influenza, (if safe in humans and we'll know soon) . Just absolutley huge I say, and with the animal rule it can (AND WILL- IF SAFE IN HUMANS) go from animal efficacy + human safety to NDA for influenza (read avian flu) outbeaks.
How many times did King mention tarvacin as a means to stop 'widespread outbreaks of viral infections' ?
His whole intro was on historic flu pandemics.
I think they've got some good flu data.
We'll see.
j
SK's Tarvacin Presentation
just a few things I jotted down that stuck out.
now, back to my wine, and someone who is dragging me away from the keyboard by my ear, and her teeth.
j
received FDA approval for P1 any cancer including; lung, pancreatic, breast, liver, others in which we demonstrated very significant pre-clinical anti-tumor activity.
Viral Therapy: Filed IND, received approval, initiating trial.
Bio-defense: tarvacin as a means to stop widespread outbreaks of viral infections.
(scary flu data stats....) control of widespread viral outbreaks
tarvacin recognizes a stable target common to all enveloped viruses.
enveloped virus examples; marburg, lassa, ebola, HIV, HCV, influenza....
shown the ability to arrest development of disease even at late stages of infection.
Tarvacin will be effective against viruses that don't even exist yet due to the MOA
for every virus we know about there are hundreds we don't know about..
(tarvacin MOA speil.......)
"AS A SUMMARY OF THE PRECLINICAL DATA THAT WE'VE GENERATED TO DATE, WE'VE FOUND THAT TARVACIN RECOGNIZES MULTIPLE ENVELOPED VIRUS STRAINS, IN FACT - EVERY ENVELOPED VIRUS STRAIN WE'VE TESTED TO DATE. THIS DATA WILL BE PRESENTED NEXT WEEK AT BIO 2005. "
ALSO SHOWN TO RECOGNIZE INFECTED CELLS FROM EVERY ENVELOPED VIRUS STRAIN.
(Lassa guinea pig info)...
"WE'VE ALSO SEEN EVEN MORE SIGNIFICANT ANTI-VIRAL EFFECTS IN OTHER ANIMAL MODELS, AND AGAIN THAT DATA WILL BE PRESENTED AT THE BIO CONFERENCE NEXT WEEK."
animals re-treated are permanently immune to further infection.
combine the animal data with human safety data from HCV & cancer trial for relatively rapid approval for widespread viral outbreaks. (animal rule).
(it really sounds to me like PPHM and the govt are doing some influeza testing, and tarvacin is in line if proven safe in humans to eventually be a frontline treatment for flu pandemics. I expect to hear influenza data at next week's conference! IMO, j)
In addition we are developing tarvacin for common viral applications. There, we've collected data on hepatitis C virus, we're continuing studies with HIV, influenza, and we're looking at some other potentials there. We will then complete human trials for each of those virus types (fast track obviously for HIV etc) "a more standard route to FDA approval"....
currently in discussions to treat viral diseases in primates, marburg, ebola....
"We have current collaborations with NIAID that are underway to screen against viral bio-terrorism and health threats. This is quite an extensive list of viruses they're testing the drug against. In fact, their indication to us was that this is one of the first times they've ever tested such a broad spectrum of viruses with a single agent."
mskso,
re:"PPHM is using one dose in the protocol, if it yields a drop of 1log in viral load and there are no harmful side effects observed with this one dose....then the FDA should let PPHM move into a Phase Ib dose escalation study fairly quickly.....it's also apparent that the safety data generated will be used to support an NDA for perhaps some biothreat indications....."
According to my understanding, PPHM is using 1 infusion per patient, but a range of doses (does escalation) in this very first study. (!)
And yes, SK came right out and said it- safety data from this study, (if it's good) and animal efficacy for biothreat viruses, (which we will apparently learn more about next week), will be used for NDA's! for lassa, marburg, etc, etc, etc,..
It could happen very fast with this animal rule. Tarvacin is the perfect thing (according to the preclinical data) to soar through the animal rule- for the broadest indications of any antiviral drug ever.
We are very close to potentially big news that would actually make news, news for average americans who have never heard of Peregrine or Tarvacin before. But of course I'm preaching to the choir here ;)
All it will take is safety data from the HCV trial... then- BAM.
j
EZ/ ms kso/ etc, re: R&R event-
I'd be happy to go. It's walking distance as I'm just a ways up Central Park West, and the R&R thing is on Central Park South at the St Regis.
Is there any cost of admission? I'd put on a hip outfit and head down as I did to UBS last fall, as long as there wasn't a big ticket to get in.
Any info would be appreciated.
Of course, I "could" simply put on my tux and walk in with a platter of shrimp...
j
ms kso- hair-trigger HAMA possibly a concern? That'd be my guess. j
Peregrine Pharmaceuticals Opens Patient Enrollment for Its Tarvacin(TM) Phase I Solid Cancer Therapy Clinical Trial
Friday June 10, 7:00 am ET
TUSTIN, Calif., June 10 /PRNewswire-FirstCall/ -- Peregrine Pharmaceuticals, Inc. (Nasdaq: PPHM - News), today announced the initiation of its Tarvacin(TM) phase I cancer therapy clinical trial. The clinical trial is open to patients with any advanced refractory solid tumor malignancy. The initial clinical centers open for patient enrollment are the Scottsdale and Tucson sites of the Arizona Cancer Center.
The clinical trial is designed to enroll up to 28 patients with advanced solid tumors that no longer respond to standard cancer treatments. The objectives of this open-label dose escalation study are to (i) determine the safety and tolerability of Tarvacin(TM) administered intravenously to patients with advanced cancer; (ii) characterize the pharmacokinetic profile of Tarvacin(TM) and; (iii) define the dose-limiting toxicities, maximum tolerated dose and/or maximum effective dose of Tarvacin(TM). Patients who demonstrate an objective response to therapy may be offered continued treatment on an extension protocol.
About Tarvacin(TM)
Anti-Phospholipid Therapy is Peregrine's novel approach to treating cancer, viral infections and certain other diseases. It is based on the finding that aminophospholipids, which are basic components of the inner surface of the cellular membrane, become exposed in certain disease states. Tarvacin(TM) is a chimeric monoclonal antibody that binds to the phospholipid, phosphatidylserine and is part of Peregrine's Anti-Phospholipid Therapy platform. Tarvacin(TM) binds directly to tumor blood vessels to inhibit growth and development of solid tumors. Tarvacin(TM) has also shown promise in the treatment of viral infections and is expected to recognize a broad spectrum of enveloped viral types. Tarvacin(TM) is currently being evaluated for the treatment of both cancer and viral diseases. Peregrine has received FDA approval to initiate two separate Phase 1 clinical trials in advanced solid cancer and chronic Hepatitis C virus indications.
About Peregrine Pharmaceuticals, Inc.
Peregrine Pharmaceuticals, Inc. is a biopharmaceutical company with a broad portfolio of products under development directed towards the treatment of cancer, viruses and other diseases. The company is in the process of initiating patient enrollment in a Tarvacin(TM) clinical trial for the treatment of all solid cancers and in a Cotara® clinical trial for the treatment of brain cancer. In addition, the company has received clearance from the FDA to initiate a Tarvacin(TM) Phase I clinical trial for the treatment of Hepatitis C virus infection, its first viral indication. Peregrine Pharmaceuticals is also developing Vascular Targeting Agents, Anti- Angiogenesis, and Vasopermeation Enhancement Agents (VEAs) for the treatment of cancer and other diseases.
Peregrine Pharmaceuticals also has in-house expertise to develop and manufacture antibodies and recombinant proteins through its wholly-owned subsidiary, Avid Bioservices, Inc., ( http://www.avidbio.com ). Avid is engaged in providing contract manufacturing services and development of biologics for biopharmaceutical and biotechnology companies, including Peregrine.
Copies of Peregrine Pharmaceuticals press releases, SEC filings, current price quotes and other valuable information for investors may be found at http://www.peregrineinc.com .
About The Arizona Cancer Center
The Arizona Cancer Center is a National Cancer Institute-designated comprehensive cancer center at the University of Arizona Health Sciences Center in Tucson and is Arizona's first comprehensive cancer center. For more information, go to www.azcc.arizona.edu.
Tally Ho ms kso!
Enjoy Kensington! I know you're a wino, but you must go to a couple of my favorite pubs in that neighborhood, the Fox and Hounds, A tiny place very near Sloane Square, (off Holbein Place if I remember correctly), and the Orange Brewery, on Pimlico road, where you can gawk at some of the finest antiques around- and drink some great beer :)
Say hi to the Duke of York's H.Q. for me,
j
EZ, that's also not really a patent. It's a PCT application, which takes years and years to "defend" in each nation/state listed (dozens), and probably will never be an enforceable patent. I know it looks like one, complete with a "patent number" etc. but it's really basically an application.
At least that's my understanding of that document.
j
OT mskso,
You might be interested in this in June's British Journal of Cancer-
"Assessment of a fragment of e-cadherin as a serum biomarker with predictive value for prostate cancer"
ms kso, the PRnewswire PR mentioned it's a dose escalation study. One dose per person, but apparently a range.
You know how fast this trial could produce data???
Follow up human viral data with a HIV IND and a PR about all enveloped viruses etc, and bingo.
mskso,
* PS- I know you work in this field and probably know most of this stuff, but once I started writing, I decided to write as if I was possibly writing to others who may nont know some of the things I was writing about.
j
Yeah, Crucell is doing great. Great company. (I'd buy some if I had the cash lately). Their cell line from a embryonic retinal cell, which is well documented in terms of low risk of contamination etc through the course of the cell line's evolution, has been the best thing available. A non-tumorigenic human cell line that expresses proteins sa bio-therapeutics... Blows away baby hamster kidney cells (BHK's) chinese hamster ovary cells (CHO's), human carcinoma lines from lung & intestine, the list goes on....
Other branches of the field (as I'm sure you know) use a few other mammalian lines from dogs, monkeys, and then there are insect cell lines, and microbial methods (e-coli, yeast etc). There is even newer technology involing plants, tobacco, and lately- duckweed to be specific.
They're all fine & dandy, and I agree that the Crucell technology is the best in terms of what's cranking out the protein drugs these days. BUT.....
THe Crucell line is not without it's concerns. The FDA is not gaga over it for a few reasons. Granted, it is the best ofd the present options, but there are issues of causing tumors in nude mice (most likely because of the particular viral vector used). The other thing is to me- the most interesting and important.
Glycosylation. "Post translational modificaitons".. Human therapuetic proteins are decorated in just the right way. If not, we recognize them as foreign, and I won't bother going on because you know all about this stuff.... Anyway, the great thing about the Multicell line is that it's a non-tumorigenic line of HEPATOCYTES, not retina cells, kidney cells, intestinal tumor cells, etc.. Hepatocytes are what MAKE all the bodies natural therapeutic proteins- the common ones are the clotting factors, antitrypsin, antithrombin, albumin, the defensins/antimicrobial peptides, etc, there are hundreds. Actually the antitrypsin types- the protease inhibitors, are the most interesting and promising IMO in terms of protein drugs. It's quite a growing branch of drug discovery these days. Multicell's hepatocyte line(s) get it right- for the first time ever. Because that's where the things really come from in the first place. Besides all the "native" proteins like the few I mentione above, the cell line can/will/is able to produce other non-native bio-therapeutics in the same way as the PER.C6 line does, by a viral vector ferrying in the genetic blueprint to make whatever it is you're looking to express, but the hepatocytes will sugar-coat it better than the retinal line, because that's what they do.
IMO, Multicell stands to becime "the next Crucell", or actually eventually pass/suplant them because of the nature of the technology. The recent PCT application needs to be read, very carefully. You will see some very interesting things in it that have not been discussed yet on any boards.
The Multicell line is well documented, no chance of any contamination issues, serum fee and all that... And it does not cause tumors in nude mice, and it glycosylates the molecules in perfect human fashion because it is the ultimate cell to do it right. I expect to see deals/collaborations in the coming months.
As for the BMY news, yeah it's great news. I hope the cell lines for drug discovery tox & ADME catch on now that BMY has jumped on board, (at three seperate sites). But I'm primarily here for the protein manufacturing possibilities.
A little farther out the speculation curve- I believe they will also have an actual proprietary therapeutic or two, made by their hepatocytes, that fills an unmet need/mkt in a big way. They will need to collaborate with a cGMP co obviously.
One last thing- as great and as versatile as the Crucell tech is, you wont see them making any of the really complex large proteins, like many of the plasma factors are, which Multicell's tech can do.
Way out- my pet theory- :) How this for down the road?
Lipitor is the biggest blockbuster right? And basically the field of cholesterol reduction is huge for pharma. If you do some reading, they're all looking into HDL's now. Rather than lowering LDL's, as the statins do, raising HDL's for a better ratio (AND actual plaque reduction) is the future. I,'m sure you remember Esperion Pharmaceuticals. They were the ones who found those special few Italian folks with the mutates ApoA1, they named ApoA1-"Milano". It showed in a small P2 study to actually reduce plaque on artery walls- first time ever for that kind of result. Pfizer bought Esperion for big bucks very quickly after that study came out. They are now working on advancing the ApoA1-Milano trials, BUT! - trials were on hold while they tried to make enough of the stuff! It is a very complex protein, obviously- it's an HDL, they're huge, hundreds of water molecules on the inside, very crazy stuff on the outside etc.. THe current recombinant method- which- I forget but I think they're using an insect line or CHO's- I need to check it's been a while. Anyway- the process to make the molecule takes over a year and requires 17 steps.
NOW- remember- HDL's are made where? By our hepatocytes, naturally. To me, as a jazz musician :), it's a no-brainer for Pfizer, who owns the "Milano"HDL variant genetic material, and also has the rights to experiment with the Multicell hepatocytes, to try to vector in the gene for the hepatocytes to start expressing this HDL, in a way that only hepatocytes could.
These types of reasons are why I'm so positive on the future of the company. The Multicell cell line can and should take a spot in the pharmaceutical manufacturing process, in a similar way to what the Crucell retinal line has done, to produce the most human therapeutic proteins yet- many that Crucell can't make, and many that it can..
The market for the "native" proteins (clotting factors, antitrypsin and OTHER natural protease inhibitors with implications for COPD wink wink) is huge as it is, and very un-met in terms of supply and demand, and the current products are either from pooled and seperated human plasma donations (not good), or recombinant metheds that don't get it right.
Neither method is keeping up with the demand.
Well, gotta start my day & get outa here,
j
wantoberich,
Thanks for bestowing the priveledges and responsibilities.
I'm honored.
I didn't realize you had sold because of the "impending" RS.
I had thought you were a regular "RB'er" with a different Ihub name. I hope your decision to sell wasn't influenced by tradenride. The company's future has never been brighter IMO.
"tradenride posted he was buying more at presplit .30's(post 1.50 - 1.75ish).
then posted he got out at pre .24 (post 1.20ish)
then posted opinions that it was likely the company would be changing the proxy the day of the vote(?!) to a higher RS ratio? LOL, jeez, I would have been very upset as I had already voted :)
(in a nut-shell, that was a silly moot argument, but I suppose it could have spread a bit of fear in some).
then he posted that if the pps was .15 pre (.75 post), he'd consider buying back in at .30 - .40 post split.
Well, it split at .245, and now sits at 1.49
(after three consecutive up days since the split).
There are several Fa2N-4 presentations with new data in the next two weeks, and obviously potential for a few other "surprises".
j
katie,
all good questions.
Yes it's a big issue for all BP, and IMO BMY is leading the way. Have a look at the past several issues of Drug Discovery and Development, or look into Stanley Hefta's group in Princeton doing cutting edge work on predicting idiosyncratic hepatotoxicity. I wasn't surprised to see tham as the fist BP client.
There is nothign I can find at the USPTO about this, and I assume they were filed around the same time. Don't know. I DO know and understand very well the advantages of the Multicell line compared to the Crucell line. Grya area is timeline, ROI :)
Re: Newmin gettign the word out. According to the company, it is just beginning. I'm expecting them to show up at UBS in New York this fall, we'll see. As for specific pharma in vitro DDI ADME tox tech conferences, there are a half dozen in the nexr few weeks for the little cell line. I think the company timed the RS as bestthey could, and so far so good- pps is up about 60% from it's post reverse split announcement low, and up about 20% since the split last week...
fingers crossed for continued momentum,
j
kso,
hey there :)
Re: Bristol Myers- they've been using the cell line for many months now, and have put out a few papers referencing the line in their work. It's not 'in the process' of being validated, except for other newer uses, and tweaking. (They've been transfect another older line of immortal human hepatocytes for some other tox related work, and it would be reasonable to think they're experimenting with the Multicell lines as well. I'd be happy to send you realted papers on the topic if you want. They're a leader in BP in terms of revamping the easrly pipeline with respect to liver toxicity issues.
Anyway- something you should be interested in is this- from an international patent application of Multicell's relating to their protein expression platform (as compared to Crucell)-
...which is THE reason I've been patient with my investment here, as I expect deals to materialize on this front.
----------------------------------------
54) Title: PROTEIN PRODUCTION SYSTEM
(57) Abstract: A method for producing competent gene products in human cells, said method comprising the following steps: Pro
viding a DNA construct in which a gene encoding a protein of interest is operably linked to a modified heat inducible promoter;
Introducing said DNA construct into a human cell line, either by transformation or by transfection to form transformed or transfected
host cell lines; Subjecting said transformed or trasnfected cell lines to a transient increase in temperature and permitting the tans
lation to protein to occur after the temperature has been returned to normal growth temperatures of the said host cells, whereby the
production of said protein of interest occurs.
PROTEIN PRODUCTION SYSTEM
The present invention relates to an Optimized Protein Production System using a Stable and
5 Competent Human Hepatocyte Cell Line
BACKGROUND
Therapeutic proteins have been known in the scientific and medical communities since the early
twentieth centuiy, but the small amounts harvestable from tissues and urine made therapeutic
replacement difficult if not impossible. In the 1 980s, advances in genomic teclmology have directly
10 facilitated the identification, isolation, and characterization of genes responsible for the production of a
great number of potential therapeutic proteins ('biotherapeutics').
Recombinant DNA technology allows the large-scale manufacture and production of many therapeutic
proteins. This approach may use either a prokaryotic or eukaryotic source of cells for propagation. The
function and efficacy of any protein - and, by proxy, a therapeutic protein, depends mainly on the gene
15 sequence; however, several post-translational modifications to the protein may also play a crucial role
in the ability of the protein to function with maximum efficacy.
Post-translational modifications (PTMs) change the property of the side groups of the amino acids -
the building blocks of proteins - such that they alter protein function. Collectively, these post-
translational modifications contribute significantly to the final structure and function of the protein.
20 Therefore, when therapeutic proteins are made for use in humans, it is thought to be important to have
the human pattern of post-translational accompaniments on the protein.
Industry example: The original production of EPO in prokaryotic cells failed because the post-
translational patterns imparted by the bacterial host-cell to its transfected human gene product were
neither correct nor sufficient to confer appropriate levels of clinical efficacy to the drug. Therefore, the
25 decision to produce EPO in mammalian - but not human - cells was an important point in the evolution
of the drug.
2
DOCUMENTED DIFFERENCES BETWEEN NATIVE VS RECOMBINANT THERAPEUTIC
PROTEINS
Reports on the importance of correct PTMs with respect to biological activity/therapeutic efficacy of
biotherapeutics abound, particularly in the view of the impending flood of new biotherapeutic
5 molecules in clinical development. Below are some examples of reported studies:
The glycosylation (a major PTM) of recombinant glycoproteins can profoundly affect their
biological activities, including circulatory clearance rate, and recombinant proteins that are
correctly glycosylated have significantly longer sermn half lives than incorrectly glycosylated
structures (Chitlaru T, Kronman C, Zeevi M, Kam M, Harel A, Ordentlich A, Velan B,
10 Shafferman A (1998). Modulation of circulatory residence of recombinant acetylcholinesterase
through biochemical or genetic manipulation of sialylation levels. Biochem J 1998 336:647-658.).
Important glossary terms:
Native protein: a protein that is routinely synthesized by a given tissue, organ, cell in the natural
physiological state, in absence of any manipulation or engineering.
15 Recombinant protein: a gene product (protein) that is obtained after genetically engineering of a cell or
organism.
Misaizu T, Matsuki 5, Strickland TW, Takeuchi M, Kobata A, Takasaki S (1995). Role of antennary
structure of N-linked sugar chains in renal handling of recombinant human erythropoietin. Blood
86:4097-4104.) found that the nature and degree of glycosylation of recombinant human
20 erythropoeitin (EPO) profoundly affected the in vivo activity. Incorrect glycosylation patterns
enhanced the total body clearance rate more than three times and resulted in a much lower activity for
stimulation of erythroid progenitor cells.
In many cases, there are significant differences in glycosylation between native and recombinant
glycoproteins, between recombinant forms expressed in different cell lines, and between related
25 glycoproteins from different organs. Landberg E, Pahisson P, Krotkiewski H, Stromqvist M, Hansson
L, Lundblad A (1997). Glycosylation of bile-salt-stimulated lipase from human milk: comparison of
native and recombinant forms. Arch Biochem Biophys 344.94-1 02.) found differences in the
glycosylation of native and recombinant forms of bile-salt-stimulated lipase (BSSL) from human milk.
Native BSSL contained a high amount of A2F family N-glycans whereas recombinant forms expressed
30 in CHO or mouse fibroblast cell lines had predominantly A2 family glycans.
3
Jacquinot PM, Leger D, Wieruszeski JM, Coddeville B,?Montreui1 J, Spik G (1994). Change in
glycosylation of chicken transferrin glycans biosynthesized during embryogenesis and primary culture
of embryo hepatocytes. Glycobiology 4:617-624.) studying the oligosaccharides of transferrins from
chicken serum, chicken embryo serum and from the culturedmedium of chicken embryo hepatocytes in
5 primary culture found each had distinct glycosylation pattems.
Tanigawara Y, Hon R, Okumura K, Tsuji J, Shimizu N, Noma 5, Suzuki J, Livingston DJ, Richards
SM, Keyes LD, et al. (1990). Pharmacokinetics in chimpanzees of recombinant human tissue-type
plasminogen activator produced in mouse Cl 27 and Chinese hamster ovary cells. Chem Pharm Bull
(Tokyo) 1990 Feb;38(2):517-22) demonstrated that two preparations of r-tPA's (recombinant tissue
10 plasminogen activators) with different carbohydrate structures showed different pharmacokinetics,
strongly suggesting that the carbohydrate structure can affect the biological activity, hence the
therapeutic efficiency, oft-PA.
WHY NATIVE PROTEINS?
Recombinant proteins that lack correct human post-translational modifications can elicit neutralizing
15 antibodies, resulting in reduced efficacy. Moreover, recomnbinantly-produced proteins are often cleared
from circulation quickly, requiring frequent injections or pegylation to extend the half-life.
"Pegylated" proteins are costly to produce and may lose some of their bioactivity, requiring higher
dosage for the same efficacy.
Conversely, native proteins from human tissues are fully human glycosylated, providing products with
20 clear activity/efficacy/safety advantages over current therapies:
Better and broader efficacy due to the presence of naturally-occurring glycan structures and subtypes
on the final protein product as well as other post-translational modifications, more closely matching
human therapeutic requirements;
Potentially fewer side effects due to lower dosage;
25 Longer half life in circulation and little allergic response due to proper PTMs;
Lower manufacturing cost due to the extraction of multiple proteins from a single cell source and
manufacturing process;
Lower regulatory barrier due to use of a single immortalized human cell line and single manufacturing
process;
4
Less frequent injections due to longer half-life of glycosylated proteins compared to non-glycosylated
recombinant proteins produced in bacteria.
Industry example: Glycogen storage disease type II (GSDII) is an autosomal recessive disorder caused
by the deficiency of the protein GAA (acid aipha-glucosidase), a glycogen-degrading lysosomal
5 enzyme.
This deficiency results in generalized deposition of lysosomal glycogen in almost all tissues of the
body and can ultimately lead to cardiac failure before the age of two years (hence GSDII is a life-
threatening condition).
Current treatment for the disease includes repairing the deficiency by injecting recombinant protein
10 into the patient, made from either cultured Chinese Hamster Ovary (CHO) cells or secreted in the milk
from rabbits that bear the transgene for the protein under a milk-specific promoter.
Both recombinant proteins produced are extremely inefficient in their uptake into and function in
targeted tissues.
The NIH (US-National Institute of Health) announces a new technology that relates to the use of
15 hepatocytes whether in culture or in vivo for the production of native human GAA.
The NIH approach is to use human hepatocytes to produce appropriate post-translational modification
of the enzyme in cells by proper glycosylation, thereby producing a superior enzyme capable of being
easily taken up and localized intracellularly in the target tissue. Once there, the enzyme digests
glycogen present in lysosomes.
20 PROTEINS NATURALLY PRODUCED BY THE LIVER
The liver is one of the most promising organs/tissues to provide producer cells with a large spectrum
potential for delivering native proteins with therapeutic interest, either as direct biological drugs or as
validated drug targets for small drug molecule development. Indeed, this organ synthesizes a host of
important proteins, including enzymes, hormones, clotting factors, and immune factors. Several
25 proteins synthesized by the liver are necessary for proper blood functioning; these include binding
proteins and albumin, which helps maintain proper blood volume. Clotting factors produced by the
liver include fibrinogen, prothrombin (Factor II), Factors VII, VIII, IX, X and von Willebrand Factor.
Acute phase proteins (APP) are another set of plasma proteins synthesized by the liver in response to
tissue damage and inflanimation associated with traumatic and/or infectious disease. Transferrin (Tf),
30 alpha-2-macroglobulin (a2M), hemopexin are just some important acute proteins.
5
(Please refer to the APPENDIX for a comprehensive list of proteins produced by hepatocytes).
[IVIF notes: in case this issue is considered a valuable avenue to explore, all potential
disease/therapeutic targets for each protein from the appendix list can be supplied]
5 LIMITATIONS OF THE NATiVE LIVER PROTEINS APPROACH
Primary hepatocytes do not proliferate, thus production of proteins from this type of cells requires a
steady supply of new cellular preparations from human liver biopsies. This would represent a clumsy
and expensive approach with many associated problems (QA, batch-to-batch variability, almost no
standardization, regulatory hassle). This issue has been solved through TGE-Corp's in-licensing
10 strategy (Multicell Technologies' unique immortalized and standardized fully competent human
hepatocytes).
The remaining limit for producing native hepatic proteins for therapeutic or other uses is obviously
determined by the set of proteins available at a decent yield from hepatic cells in culture. The
APPENDIX section lists all major proteins that could potentially be manufactured under the label of
15 'Native Proteins'.
Any other therapeutic protein candidate will have to be produced using genetic engineering strategies.
However, even in this case, the human nature/origin of these cellular substrates (MCT's hepatic cell
lines) should guarantee the best post-translational modification process currently available, thus
leading to recombinant end-products with clear competitive advantages, including a favourable
20 regulatory outlook.
6
APPENDIX 1
BASIS FOR THE CONSTRUCTION OF HIGH EFFICIENCY, INDUCIBLE PROMOTER - GENE
CONSTRUCTS, EXPRESSING THERAPEUTIC PROTEINS.
5 An expression vector will be constructed which allows for the selection of stable transfectants by
selection for the zeocin antibiotic (Cayla) in both prokaryotic and eukaryotic cells. The Zeocin
resistance gene will be obtained as a restriction digest fragment from the pZeoSV plasmid (Invitrogen)
and will be ligated to a fragment containing a bacterial origin of replication obtained by PCR
amplification from pUC19 (New England Biolabs). This ligation mixture will then be used to
10 transform competent E. coli cells and the presence of the desired recombinant plasmid (pUC-Zeo) will
be selected for on Zeocin-containing bacterial plates. A synthetic poly(A) sequence will be obtained
as a restriction fragment from a digest of pGL3-Basic (Promega) and will be ligated into pUC-Zeo
upstream of the HSP7OB promoter and the desired recombinant (pUC-ZeoA) will selected for Zeocin
resistance. The HSP7OB driven expression cassette (Hi-Hot) will be obtained as a restriction fragment
15 from the PCR amplification of pHi&Hot-MCS (V3) (David Harris, University of Arizona) from which
an XhoI fragment in the multiple cloning region has been deleted. The Hi-Hot expression cassette will
be ligated into pUC-ZeoA downstream of the synthetic poly(A) sequence and the desired recombinant
(pHiHot-Zeo) will be selected for Zeocin resistance. Genes to be expressed under the control of the
Hi-Hot system can be inserted into the unique XhoI and XbaI sites derived from the multiple cloning
20 sequence of pHi&Hot-MCS. The Hi-Hot plasmid constructs are derived from those in Tsang et al.,
Biotechniques, 20:51-52, 1996 and Tsang et al., Biotechniques, 22:68.
In a comparison of expression systems in transient transfection experiments using the Lewis Lung
Carcinoma cell line expressing Interleukin-2, the following results were obtained:
25
Hsp7OB promoter - 468 pg/ml of interleukin-2
CMV promoter - 573 pglml of interleukin-2
Hi-Hot promoter - 18,409 pg/ml of interleukin-2
30 THE HI-HOT PROMOTER WAS 39.3 TIMES STRONGER THAN THE HSP7OB
PROMOTER AND 32.1 TIMES STRONGER THAN THE CMV PROMOTER
14
APPENDIX 3
EXISTING hUMAN CELL LINE PRODUCTION SYSTEMS TODAY
The initial success of biotechnology has been driven some 30 years ago, by the ability to transfer DNA
sequences into living cells and make them produce therapeutic proteins/peptides on a large scale. The
5 production of recombinant insulin was the first example of such a process, achieved in the l97Oies by
introducing the insulin gene into bacteria, the simplest cell type available. These 'engineered bacteria'
then produced insulin which to date is still successfully used to combat diabetes.
As the biotechnology industry became interested in the production of more complex protein drugs,
more sophisticated producer cell types were required as a production platform. In the early 8Oies, the
10 large-scale manufacturing of therapeutic proteins in mammalian cells represented a major
breakthrough. To date, animal cells, such as rodent cell lines, and in particular the Chinese Hamster
Ovary cell line (CHO), represent the most important platfonn for the production of
biopharmaceuticals, including some blockbuster biotech drugs.
However, in recent past clinical trials, in particular with recombinant murine antibodies, it rapidly
15 became clear that nonhuman antibodies have the potential to elicit an immune response, thus blocking
the efficacy of the treatment. These observations underscored the now recognized importance of
correct 'human-type' post-translational modifications of complex protein products.
PER.C6TM
The company Introgen b.v. (Leiden, NE), now Crucell, took the bioproduction process one step further
20 from rodent to a human production platform. PER.C6TM is an expression platform that consists of a
human cell line that can produce biophannaceuticals for human therapeutic use. The PER.C6 cell line
was generated from human retina-derived primary cells, which were immortalized by insertion of the
adenovirus El gene.The cell line is derived from a single source of healthy human cells in a controlled
and fully documented manner. The company has immortalized the cell so that it can replicate itself
25 indefinitely, unlike normal human cells, a prerequisite feature essential to the production of
recombinant biopharmaceutical products in sufficient quantities for commercial distribution.
Currently, Crucell promotes its PER.C6 production platform for the:
Production of monoclonal antibodies through rDNA technologies;
Production of various therapeutic proteins other than mAb's;
30 Vaccine production
15
Adenoviral vector production for gene therapy applications
Functional genoinics
WHY PER.C6TM
Crucell, the PER.C6 owner company, claims its platform to represent today's industry standard for
5 applications such as 1 - 4 above. The main reason to this stems from the following assertion:
"Crucell's Technology Maintains 'Human' Glycosylation Patterns."
Major advantages of using PER.C6 cells for biopharinaceutical production:
Post-translational modifications (in particular Glycosylation): Optimal recombinant therapeutic protein
products, in terms of half-life, biological activity and immunocompatibility contain human glycan
10 structures (i.e. glycosylation patterns). Mammalian cells like CHO (the biotech 'workhorse') or other
established non-human animal cell lines add non-human glycan structures to recombinant proteins or
antibodies. PER.C6 cells perform human glycosylation patterns, resulting in higher biological activity
and longer half-life.
Regulatory issues: As PER.C6 has been developed as a manufacturing platform for
15 biopharmaceuticals, extensive documentation concerning the generation and characterization of the
cell line has been assembled from the start. This documentation has been deposited as a biologics
master file (BMF) with the FDA. PER.C6 has been approved for the generation of recombinant
adenovirus for gene therapy trials, and has been accepted for Phase I/LI clinical trials of an HIV
vaccine being administered to both healthy and immnunocompromised individuals.
20 Production yields - monoclonals: Currently used expression platforms for the production of
monoclonal antibodies are CHO and NSIO cells, with average expression yields amounting to
approximately 0.5 g/l in final production processes. PER.C6 produce similar levels of antibodies in a
non-optimized system and is expected to produce significantly more in optimized fed batch culture
systems.
25 Intellectual Property: Crucell, Leiden, NL, wholly owns the technology and know-how associated with
PER.C6, which translates into a transparent patent situation, which is far from being the case for all
other production platforms for biopharmnaceuticals.
Flexibility of use: The cells grow readily as adherent or suspension cultures, in serum-free and animal-
component-free culture systems and can be easily transferred from one medium or growth condition to
30 another.
16
Scalability: The presence of the adenovirus El gene inhibits apoptosis of PER.C6 cells, resulting in
high viabilities when grown in batch production cultures. PER.C6 cells are easily scalable - the cells
are currently grown in 2,500 L reactors and further upscaling is in progress.
Stability: Transfection of PER.C6 cells with expression plasmids is efficient, as is subsequent
5 generation of stable sub-clones. Importantly, high expression levels of recombinant proteins are
observed in the absence of gene amplification, giving a considerable time advantage over the use of
cell lines that require amplification for efficient protein expression.
In a May 2002 hearing (http://www.fda.gov/obrrns/dockets/ac/Ol/briefing/375Obl_01 .htm), the FDA
considered the potential risks in using two novel cell substrates, i.e. HEK293 cells and PER.C6 cells.
10 These cell lines were developed by transforming human embryonic kidney cells (293) and human
embryonic retinal cells (PER.C6) with the transforming early region 1 (El) of adenovirus type 5
(Ad5). Since cell lines such as 293 and PER.C6 express the AdS El region, they are able to
complement the growth of defective AdS vectors which have been 'crippled" by deletion of El.
Defective Ad5 vectors have been engineered to express foreign genes such as those from human
15 immunodeficiency virus (HIV), the causative agent of AIDS, and vectors of this type are thought to
have significant potential for vaccine development because of their demonstrated ability to generate
cell-mediated immune responses to HIV. However, a feature of regulatory importance associated with
AdS-transformed cells is their capacity to form tumors in immunodeficient animals such as nude mice.
In considering potential risks associated with the use of so-called Designer Cell Substrates - i.e.,
20 neoplastic cells derived from normal cells transformed by defined viral or cellular oncogenes or by
immortalizing cellular genes (e.g., telomerase) - OVRRJCBER is considering the approach outlined
within the framework of a "defined-risks" assessment Lewis et al., "A defined-risks approach to the
regulatory assessment of the use of neoplastic cells as substrates for viral vaccine manufacture", In
Evolving Scientific and Regulatory Perspectives on Cell Substrates for Vaccine Development. Brown,
25 Lewis, Peden, Krause (eds.) Develop. Biol. Stand.. This framework is intended to examine, and
wherever possible, to quantifr the potential risk of "transmitting" the tumorigenic components of the
cell substrate used for vaccine production, and determine whether that "transmission" might pose a
risk, particularly an oncogenic risk, to vaccinees. Factors that could influence the risk associated with
the use of Designer Cell Substrates include (a) the known mechanism of cell transformation leading to
30 the development of tumorigenic cells; (b) residual cell substrate DNA; and (c) the presence of
adventitious agents, especially oncogenic viruses.
17
CRUCELL
Crucell discovers and develops fully human biopharmaceuticals that utilize the immune system to
combat disease. Crucell's proprietary technology platforms, MAbstractTM , AdVacTM and PER.C6TM,
enable the discovemy, development and production of novel antigens, Antibodies and Vaccines. Crucell
5 offers its technology to pharmaceutical and biotechnology partners, and utilizes them to create
Crucell's own product pipeline.
PER.C6TM is a human cell manufacturing platform, which has become the industry standard for
production of recombinant adenoviral vectors. PER.C6TM has also proven to be a superior platform for
the production of antibodies and vaccines.
Crucell has 19 licensees for its PER.C6TM technology, including Novartis, Pfizer, GSK, Aventis,
Genzyme and Schering.
PER.C6TM in Crucell's Press Releases
PER.C6TM is a human cell platform for the development and manufacturing of bio-pharmaceutical
products such as antibodies, proteins and vaccines. The superior yields and scalability of PER.C6, as
well as the extensive history and safety documentation render PER.C6 the safe, cost effective and
large-volume manufacturing platform that the pharmaceutical industry requires. Having launched the
use of PER.C6 as a vaccine platform through an exclusive licensing agreement with Merck & Co. for
their HIV vaccine, Crucell aims to expand its PER.C6 business in the field of vaccines. The current
agreement with Rhein Biotech endorses this strategy.
18
CLAIMS
1. A method for producing competent gene products in human cells, said method comprising the
following steps:
Providing a DNA construct in which a gene encoding a protein of interest is operably linked to a
modified heat-inducible promoter,
Introducing said DNA construct into a human cell line, either by transformation or by transfection to
form transformed or transfected host cell lines,
Subjecting said transformed or trasnfected cell lines to a transient increase in temperature and
permitting the translation to protein to occur after the temperature has been returned to normal growth
temperatures of the said host cells, whereby the production of said protein of interest occurs
2. The method of claim one in which the modified heat inducible promoter is the Hi-Hot promoter.
3. The method of claim one where the human cell line is a competent human hepatocyte cell line.
4. The method of claim 3 where the gene expressed encodes a therapeutic protein.
5. The method of claim 3 where the gene expressed includes therepeutic genes such as interferons,
interleukins, blood clotting factors, insulins, growth hormone, urokinase, EPO, TPA, FSH,
somatostatin, antibodies, DNAase, myoglobin, pro- and anti-angiogenesis factors and proteins of
veterinary interest.
6. The method of claim 3 where the gene expressed is a natural liver protein.
-------------------------------------------------------------
APPENDIX 2
SHORTLIST OF NATIVE IIEPATIC PROTEINS
Abbreviated
Protein Name(s)
202 lii 202
3alpha p a iy roxysterol 1 y ro 10 e y rogenase, 1 C9, aldo-keto reductase
HSD/DD 1C9
A2-Mag a p -macrag o u in
A2 UG alpha-2u globulin clone RAO 01
A2UG2O7n alpha-2u globulin 207
AA1 aiphal-antitrypsin
ACC acetyl-CoA carboxylase
ACE angiotensin converting enzyme , DCP, DCP 1, dipeptidyl carboxypeptidase 1,
angiotensin I converting enzyme, peptidyl-dipeptidase A 1, CD 143, dipeptydyl
peptidase, Kininase II, ACE 1
ACO acyl CoA oxidase
ACS acyl-coenzyme A synthetase, long-chain-acyl-CoA synthetase
ADB aldolase B
ADHi alcohol dehydrogenase 1, class I, alcohol dehydrogenase alpha subunit
ADH2 alcohol dehydrogenase 2, class I
ADH3 alcohol dehydrogenase 3, class I
AFP alpha-tetoprotem
aG aiphal-globin
AGG Agamma globin, hemoglobin gamma chain
AGP alpha-i-acid glycoprotein
ALAD delta-aminolevulinate dehydratase, porphobilinogen synthase
ALB albumin
ALDH-3 Class 3 aldehyde dehydrogenase, fatty aldehyde dehydrogenase, aldehyde
dehydrogenase microsomal
alpha 113 alpha 1 -inhibitor 111
aIpha2MRILRP alpha2-inacroglobulin receptor/hpoprotein receptor-related protein
AMIl acute myeloid leukemia 1, CBFA2, core-binding factor, runt domain, alpha subunit
2
ANIG angiotensinogen
ERSATZBLATT (REGEL 26)
WO 2004/111246PCT/CH2004/000363
8
Protein Name(s)
7OX acyl CoA oxidase , acyl-CoA: oxygen 2-oxidoreductase
apolipoprotein(a)
apoAl apolipoprotem Al
ApoA-I apolipoprotem A-I
apoAlI apolipoprotein All
ApoA-I V apohpoprotein A-I V
apoB apolipoprotein B
apoC-Ilt apo ipoprotein -
ApoD apohpoprotein D
apoE apohpoprotein B
apoVLDLII very low density lipoprotein II, apovitellenin I
AI(G arginase
AT1a-R angiotensin II type lA receptor, angiotensin receptor (All)
ATPlA1 Nal(-AlPase alpha 1 subumt, NAKA alpha 1
beta2-AR beta2-adrenergic receptor
beta-IF N virus induced interferon-beta, IFN beta
BI complement factor B
BFIBR tibrinogen B-beta subunit
BGP biliary glycoprotein, C-CAM-i, cell CAM-l, BGP-l, BUPi, antigen CD66,
CD66A antigen
C/BBPalpha CCAAT/enhancer binding protein alpha
C3 complement C3
C4BP C4b-bmndmg protein
cAspAl cytosohc aspartate aminotransterase
catalase catalase, CAl, CAS 1
Cdc 7 Cell division cycle 7
CEBPA CCAAI/enhancer binding protein alpha
CEIP cholesteryl ester transter protein
COLlAl collagen, type I, alpha 1
CItBPII cellular retinol-binding protein II
CRP C-reactive protein
CSP 1 carbamoylphosphate synthetase 1
Cypi lA cytochrome P450 cholesterol side chain cleavage , P45Oscc
ERSATZBLATT (REGEL 26)
Abbreviated
WO 2004/111246PCT/CH2004/000363
9
Protein Name(s)
cytoebrome P-450 l7alpha hydroxylase/C1'/-20 lyase , P450c1 I
Cypl9 aromatase, cytochrome P450XIXA1 , estrogen synthetase , P450arom
CYPlAl cytochrome P450 tamily IA 1, P1-450, cytochrome P-4501A1
Cyp2IWf cytochrome P450 steroid 21-hydroxylase , P450C21 , CypXXIA1 , 21-OHase
CYP27 Cytoebrome P-450c27
CYP2B2 cytochroine P4501132
CYP2C12 E1ieP43ThiiiiIF2Tl2
YP7 cholesterol 7-aipha-hydroxylase , cholesterol 7-aipha-monooxygenase, cytochrome
P450 VII
DRA MHC class 11 HLA-DRA
ei24 etoposide induced protein 2.4, PIG8
EKEF erythroid Kruppel-hke factor
Epo erythropoietin
FlO coagulation factor X, coagulation tactor 10, stuart tactor, 110
F7 tactor VII, vitamin K-dependent coagulation protein
coagulation factor VIII, F8C, procoagulant component, antihemophihc tactor, AHF
F9 coagulation factor IX , Factor IX, Factor 9, christmas factor, FIX, 19
lAS fatty acid synthase
TI7tor Fas ligand receptor, Fas antigen, Fas death receptor, APp-i antigen, CD95 antigen,
TNFRSF6, APTi, FAS
FCH ferrochelatase, protoheine terro-lyase, heme synthetase
FGG FIBRINOGEN GAMMA, FIBG
librinogen gamma-tibrinogen submt
gamma
FPPS tarnesyl diphosphate synthase , geranyl-diphosphate: isopentenyl-diphosphate
geranyltranstransferase
FIi-I territin, heavy chain
G6Pase glucose-6-phosphatase
GAD 1 glutamic acid decarboxylase 2, 67k1)a glutamate decarboxylase , GAD67
GAlA-i GAiA-l
Gbp- 1 guanylate-bmding protein- 1, mGBP-1, mag-1, macrophage activation -1
Gbp-2 guanylate-bmding protein-2, mGBP-2, macrophage activator
GUG Ogamma-globin
ERSATZBLATT (REGEL 26)
Abbreviated
WO 2004/111246PCT/CH2004/000363
10
Protein Name(s)
GI( glucokinase, hexokinase D, hexokinase type IV, 1-1K4, AlP:D-hexose 6-
phosphotransferase
GLOB-A alphal-globm, hemoglobin aiphal chain
GLOB-B beta-globin, hemoglobin beta chain
GLOB-BM betaM-globm, hemoglobin beta major chain
GLOB-Z zeta2-globm, hemoglobin z2 chain
GPA glycophorin A
GPAT mitochoncirial glycerol-3 -phosphate acyltransferase
GPC glycophorin C
gpD glycoprotein D, major subunit ot Dutty blood group
GSHPx cytosohc glutathione peroxidase, cellular glutathione peroxidase, GPXT
T2 Glutathione S-transferase A2
GST-P glutathione transtrase P , placental glutathione transterase, glutatmon S-alkyl
transferase
GSTYA glutathione S-transterase Y a subunit
STY glutathione S-transterase Y a subunit
H-2Dd MHC class I H-2Dd
H-2Kb MHC class I H-2Kb
H-2Kk MHC class I H-2Kk
12T ]VIHC class I H-2Ld
HCEH hepatic neutral cholesterol ester hydrolase , neutral cholesterol ester hydrolase
hepatic cholesterol ester hydrolase
RD hydratase-dehydrogenase , enoyl-CoA hydratase /3-hydroxyacyl-CoA
dehydrogenase , bifunctional enzyme
HG haptoglo bin
RIG hexokinase 3, HK III
TGW RMG-CoA reductase, 3-hydroxy-3-methylglutaryl-coenzyme A reductase , 3-
hydroxy-3-methylglutaryl-coA reductase
HMGCS 3-hydroxy-3-methylglutaryl coenzyme A synthase , hydroxymethylglutaryl-CoA
synthase, cytoplasmic, HMG-CoA synthase
HMGR 3-hydroxy-3-methylglutaryl coenzyine A reductase
HNF-1 hepatocyte nuclear factor-i, LFB1, LFB 1/HNF1, HNF-ialpha
HNF-i beta hepatocyte nuclear factor-i beta, vHNF-i
ERSATZBLATT (REGEL 26)
Abbreviated
WO 2004/111246PCT/CH2004/000363
11
Protein Name(s)
Hnt-3alpha hepatocyte nuclear tactor 3alpha
HNF3beta hepatocyte nuclear tactor 3 beta
HNF3gamma hepatocyte nuder tactor 3 gamma
T14 hepatocyte nuclear tactor 4
1-10-i heme oxygenase 1, hsp32
FG haptoglobin
Hpx hemopexin
hsc70 72 kd heat shock cognate
hsc7T heat shock cognate 73, HSC727P2, HSC7O, peroxisome proliferator binding
protein
flFT heat shock factor 1, HSTF1
FS heat shock factor 2, HS1F2
TPI3 heat shock protein 105
hsp25 heat shock protein 25
84 kDa heat shock protein, HSP9OB
TI2 Basement membrane-specific heparan sultate proteoglycan core protein , perlecan,
HSPG, PLC
H'1GL hepatic triglyceride lipase , triacylglycerol lipase
T1fT intercellular adhesion molecule-i , major group rhinovirus receptor, HI{V, (JD347
11-N -alpha interteron-alpha
TGFT insulin-like growth factor binding protein-i
- insulin-like growth factor 11
iNUS inducible nitric oxide synthase, N0S, hiNUS, inducible NOS, inacrophage-type
NOS, iNOS, NOS, type II, NOS2A, hepatocyte OS, hep-NOS
iP-iO intiammatory protein iO-kDa, Crg-2, gamma-lPiO, C7, INPiG, 11-hO, SCYBiO
JR insulin receptor
IRE-i interferon regulatory factor i
ltgb5 integrin beta 5
TP17 lipoprotein lipase
MCAD medium-chain acyl-CoA dehydrogenase, acyl-CoA dehydrogenase, medium-chain
specific, 2,3-oxidoreductase , ACADM
MDRi multidrug resistance , PGY 1, P-glycoprotein
mdri b multidrug resistance 1 b, P-glycoprotem , mdri
ERSATZBLATT (REGEL 26)
Abbreviated
WO 2004/111246PCT/CH2004/000363
12
Protein Name(s)
malic enzyme , malate oxidoreductase
MHMGCS mitochoncinal 3-hydroxy-3-methylglutaryl coenzyme A synthase , mitochondrial
HMG-CoA synthase
MT-I metallothionein-1
MTP microsomal triglyceride transter protein
fX1T Max Interactor 1
FE2 nuclear tactor erythroid 2, nuclear factor (erythroid-derived 2), 45kD, P45 NF-E2,
leucine zipper protein NF-E2, NFE2, p45, NF-E2
PY neuropeptide Y
TYT NAD(P)H:quinone oxidoreductase, Dl diaphorase, NMO1, NMOI{, QR, QAG
GAS 2-5' oligoadenylate synthetase, 2'-5 A synthetase
ODC ornithine decarboxylase , L-ormthine carboxilase
omithine transcarbamylase, L-ornithine carbamoyltransterase
p44 hepatitis-C-associated microtubular aggregate protein
P450c cytochrome P450 tamily I A 1
p53 tumor suppressor
1fl Phenylalanine hydroxylase
PBGD porphobilinogen deaminase, porphobilinogen ammonia-lyase (polymerizing),
hydroxymethylbilane synthase
PCNA proliteratmg cell nucleolar antigen p120
EPK phosphoenolpyruvate carboxykinase cytosohc, phosphopyruvate carboxylase
PFIF 6-phosphofructo-2-kmase/fructose-2,6-biphosphatase , 6PF-2-KJFru-2,6-P2ASE,
PFK-2IFBPase-2, RH2K
POKT1 phosphoglycerate kinase 1 somatic-type
pIgR polymeric immunoglobuhn receptor, polyimmunoglobulin receptor, polymeric IgA
receptor
pk L L-type pyruvate kinase
PPAR gamma peroxisome proliterator activated receptor gamma, PPAR gamma 3, PPAR gamma
1
PRLR prolactin receptor
Pt prothrombin, coagulation tactor II, F2
PY1T M-type pyruvate kinase, PI(M2, Pyruvate kinase, M
I{AR-beta retinoic acid receptor beta, Retinoic Acid Receptor beta 2 , RAR-beta
ERSATZBLATT (REGEL 26)
Abbreviated
WO 2004/111246PCT/CH2004/000363
13
Protein Name(s)
RB'1'N2 Rhombotin 2, T-cell translocation 2, T1G-2
514 514 product
SAA serum amyloid A
SAA 1 serum amyloici Al
SAA3 serum amyloid A3
SAAg9 serum amyloid Ag9
sgk Serut'1lireonine protein kinase
ShiP signal-regulatory protein
S(JDl Cu/Zn superoxide dismutase
Spi 2.1 serine protease inhibitor 2.1, serpin
Spi Z3 serine protease inhibitor 2.3, SPI, serpin, contrapsin-like protease inhibitor
precursor, kallikrein-binding protein, GHR-p63
SPTB beta-spectrin, SP 1131
SREBP-2 sterol regulatory element-binding protein-2
SS squalene synthase, tarnesyl-diphosphate: tarnesyl-diphosphate tarnesyltransterase
Stat6 lnterleukin-4-induced transcription tactor Stat6, IL-4 Stat
T7ET7 SCL, 1CL5
IAl tyrosine aminotransterase
'IF transterrin
Il-IPO thrombopoietin, c-Mpl ligand, megakaryocyte colony stimulating tactor,
megakaryocyte growth and development factor
10 tryptophan oxygenase
'1 0P3 DNA topoisomerase 111 alpha, TOP3A, hTOP3
TTR"" transthyretin, prealbumin
VIT-A2 vitellogenin A2
VhF-B 1 vitellogenrn B 1
V IT-Il vitellogenin hi
vWF von Willebrand tactor
XOR xanthine oxidoreductase, XDH, XDHA
zg zeta-gb bin, hemoglobin z-chain
wantobe, thanks for the offer, but I'm not interested in the responsibility. - just looking for a temporary forum while RB is unavailable switching over. I didn't realize this board was so quiet and that you have been "out" of MUCL/MCET. Out of curiosity, now that the split is done, will you be looking at a re-entry?
j
time to start using this board again? eom
wantoberich, I'm not sure what your job-offer entails, but I don't think I have the time to have any real duties/responsibilities with a forum like this.
I appreciate reading the opinions here, and I sure do appreciate this board being free of whining brats.
Hey, whadya know, looking like .45 is headed our way.
This may be my last post of the day- if that 3 posts for free rule applies.
j
delphi23,
feel free to repost, that's fine, but understand I'm no scientist, just an interested layman. There are some others on the RB board with a deeper knowledge of the science than myself- thaliomiles understands much more than I. We've done lunch here in New York a couple times and his grasp on molecular & cellular bio is very impresssive. Also- p_g_s, especially when it comes to the MUCL cell lines (since he actually worked on them with Jin Lui at Multicell).
Today's bean is from Rawanda, (Masaka).
I'm a home roaser. Once you start home roasting, there's no goin' back...
j
hey all. hi trade, so this is where you've gone to. eom, j