X-Message-Number: 25134 From: "Basie" <> Subject: Freeze-Dried Miracle Date: Mon, 22 Nov 2004 19:49:53 -0500 Maybe CI and Alcor can convince the military that soldiers will be more effective if they had a "suspension" benefit in their contract. Basie Freeze-Dried Miracle After more than a decade of developing and refining a patented process for freeze-drying and sterilizing blood platelets, an ECU researcher is on the verge of making a life-saving technology a reality . by Garnet Bass Pathology professor Arthur P. Bode can hold the symbol of $10 million and 12 years of painstaking research in the palm of his hand: a vial of white powder that looks no more glamorous than Epsom salts. But, a few years from now, in the hands of an Army medic on the operating room, that little vial could save a life. Cheryl Binkley, research assistant, examines digitized black-and-white images of StasiXT platelets photographedon a fluorescence microscope and processed with a CCD camera. Or many lives. Dubbed StasiX, the powder consists of human blood platelets freeze-dried and sterilized though a process Bode developed with collaborators at UNC-Chapel Hill. In laboratory and animal tests, the freeze-dried platelets have shown the ability to quickly stop bleeding in cases of severe injury. Best of all, they can be stored safely at room temperature up to five years - compared with a limit of five days on blood bank platelets cal for situations beyond the reach of traditional blood banks. These properties represent the potential to save the lives of thousands of people worldwide who bleed to death each year. To turn that potential into reality, Bode and his partners have formed Hemocellular Therapeutics Inc., a private company that has licensed the technology from the universities and secured about $1 million in private seed capital to refine the freeze-drying process further. Within the next few months, they plan to submit to the U.S. Food and Drug Administration a proposal to begin clinical trials in humans. If all goes according to plan, freeze-dried platelets could be in widespread use within the next few years. Bode, the lab scientist, finds himself bemused by his new corporate world with its strange language - Vertically integrated business plan? High-end supply? - and fast pace. Graduate school doesn't prepare one for this," he said. Dr. Arthur Bode removes a small vial of StasiXT from a lab freezer. In other ways, graduate school prepared him for precisely this point. The road would not be direct, however. The years in between would be occupied with patient research, beset by roadblocks and blessed by serendipity. The first seeds for Hemocellular were planted 20 years ago at UNC-CH where the pathology department was headed by a man Bode calls "one of the godfathers of coagulation and platelet science," the late Dr. Kenneth Brinkhous. In the early 1960s, Brinkhous had developed a diagnostic test for the common bleeding disorder, von Willebrands disease. The test used platelets, and given how frequently it was performed, it used a lot of platelets. To eliminate the need for a constant supply of fresh platelets, Brinkhous and his associate, Dr. Marjorie S. Read, developed a simple process for freeze-drying platelets and obtained a patent on it in the early 1980s. "No one thought of using that freeze-drying for any other use except diagnostic testing," said Bode, who first heard about Brinkhous and Read's process as a doctoral student in Chapel Hill. By 1986, Bode had moved to ECU and was directing research to prolong the shelf life of fresh platelets held in blood banks. Unfortunately, that research came to a screeching halt soon thereafter when the Food and Drug Administration declared that it would not allow platelets, which must be stored at room temperature, to be kept for more than five days because of potential bacterial growth. With the FDA announcement, Bode's funding evaporated. Looking for a new avenue of research, he talked with Cmdr. Lyn Yaffe in the Office of Naval Research. Binkley carefully positions a segment of arterial vessel with adherent StasiXT platelets for examination under the fluorescence microscope. "He said if you guys could ever learn how to freeze-dry those cells, we'd be in business," Bode recalled. "Lo and behold, the connection is made. I went running back to Dr. Brinkhous. I said you guys freeze-dry your platelets, don't you? He said yes, of course. I said, well, have you ever thought about putting those back into a human being and see if they stop bleeding? I can't believe I surprised somebody who was one of the top three people in this field, but he just looked at me and said no, we never thought of that. So he and Marjorie Read and I sat down and had a good discussion." For the military, freeze-dried platelets represented the potential to keep soldiers and sailors from bleeding to death in the midst of battle. Platelets have several properties, the most critical being their role as the body's first-responder to injury. When they sense a wound, they stick to it and promote clotting. Without platelets, chances of stopping severe bleeding are slim. Yet platelets have been missing in action from the battlefront. Currently, blood is separated into three products for blood banking. Plasma, the fluid part, can be freeze-dried and stored for years. Red cells, which carry oxygen, can be kept refrigerated for seven weeks. Refrigerated platelets, on the other hand, refuse to recirculate. At room temperature, the short shelf life of platelets renders them useless for military field operations. Civilian blood banks also run into problems. Occasionally, they run short, but overall, they discard about 20 percent of platelets, unused, when they pass the expiration date. Clearly, freeze-dried platelets - easy to store and transport - would be a major benefit. Previous attempts to produce freeze-dried platelets for transfusion date back to the 1950s. None succeeded. Nor was Brinkhous and Read's process, while adequate for diagnostics, suitable for this purpose. It was far too harsh. To maintain their clotting properties, platelets had to retain their delicate sponge-like texture and the surface "whiskers" that locate and cling to wounds. But it did present a foundation for new investigations. In 1989, with funding from the Navy, Bode and Read took on the challenge. One after another, they tried different fixatives and treatments - microwaving, air-drying, freeze-drying, a lot of "kitchen chemistry," as Bode described it. "It took about four years to get to where we could, very nicely, with fine tuning, get the platelets fixed so that we rigidified their surface no more than you had to, to be able to freeze-dry them and still have a fairly active cell once you added the water back." Eventually, tests would show that freeze-drying, or lyophilization, not only preserves the platelets' clotting properties, but enhances them. When transfused, blood bank platelets can take up to 24 hours to regain full function. Freeze-dried platelets, reconstituted in a saline solution, instantly go to work to stop bleeding. The process is amazingly simple. "There's nothing fancy about it," Bode said. "Just take a bottle [of freeze-dried platelets], shoot it into the salt solution, and swirl, swirl, swirl. It bursts back into cells." With that burst, the reconstituted platelets are ready for a bleeding patient. So far, the platelets have shown no deterioration from being stored up to five years. The only apparent use for which blood bank platelets remain superior is as preventative therapy. Blood bank platelets are preferable if the goal is to fortify a patient's blood to prevent bleeding; but if the patient is already bleeding, freeze-dried platelets promise a better solution. Above left, fluorescent antibodies illuminate the periphery of reconstituted freeze-dried platelets that have become activated at the site of a blood vessel injury. Above right, an image made with stacked optical frames in a confocal laser-scanning microscope shows that lyophilized platelets (stained red) stick appropriately to fresh platelets (stained green) at an injury site. By the mid 1990s, the project was progressing. UNC and ECU jointly patented the process and licensed it to a company called Centeon, which already produced freeze-dried plasma. The Navy continued to fund both the universities and Centeon. Though the relation- ship between the researchers and the company strained at times, they made progress on refining the process. Clinical trials, necessary to prove safety and efficacy, appeared to be only a couple of years away. Then a problem with an unrelated Centeon product ricocheted, affecting other operations. The company withdrew from the project in 1998 and returned the remaining grant money to the Navy. Bode and Read persevered and formed a tentative partnership with the Red Cross as a step toward clinical trials. The Navy reduced its funding and the researchers rewrote their timeline accordingly. Then came what appeared to be the crushing blow: Concerned that without Centeon, the product would never reach the marketplace, the Navy halted funding altogether in 2000. "Quite literally, we were within a couple of months of doing a critical study that would have established us as a viable entity for pro ducing something they desperately needed," Bode said, "and it vaporized right before our eyes." Nonetheless, one fortuitous development came from the Centeon relationship. Company officials had questioned what effect the freeze- drying process had on viruses. They had recognized what the researchers initially did not: the growing concern about the transmission of viral diseases through blood products. Soon, tests confirmed they had a product that not only fixed platelets but also sterilized them. Richard Hodges, research assistant, sets up the new cell separation equipment used to prepare platelets for the freeze-drying process. "That's the real kicker," Bode said. "This same process kills, so far as we know, all viruses and all bacteria that you can load into a platelet suspension. That means it kills the hepatitis virus. That's a terrible risk for transfusion medicine. It kills HIV. It kills the viruses that we put in there just as models of what viruses look like. So most likely, this process is going to sterilize against everything. We can't prove that, but for all the test viruses that have been tried, they show a total kill. No virus survived." The two years after Centeon pulled out dragged. Eventually, UNC-CH, which had taken the lead on technology transfer, recovered the license. Read retired. Into her place stepped another UNC-CH scientist, Dr. Tom Fischer, but no new corporate suitor came forward with an interest in commercializing freeze-dried platelets. "It became apparent to us that to get this off the ground and get it to clinical trials, we would have to do it ourselves," Fischer said. Bode demurred. "Actually, he [Fischer] is the guy who jumped into the business end of this and pulled me in with him," he said. Fischer enrolled in a class offered by the UNC-CH Kenan-Flagler School of Business. Targeted to university researchers, the course walked him through the fundamentals of how to launch a new company. With Bode's help, he did the market research and put together a business plan. Each student in Fischer's class presented his plan to a group of business, financial and legal experts the school had gathered to provide feedback. Sitting in that audience was E.S. "Stan" Eskridge, an entrepreneur who had a long history with business startups. He had launched his first business in the late 1970s. Called the Aviation Group, it became the overnight air force for United Parcel Service. When Fischer made his presentation, Eskridge liked what he heard. "I had an instinctive reaction to Tom and the technology, and of things in that course, it looked to me like this had the shortest path to revenue," Eskridge said. They talked outside of class, then met with Bode. "Fischer and Bode are both outstanding as scientists and are good people," Eskridge said. "I was convinced this particular transaction had the potential to become a poster child for how we in business, government and universities could work together." It didn't hurt that Bode and Fischer remained well ahead of any potential competition. Two other groups were known to be investigating different processes for freeze-drying platelets. Neither had achieved a similar level of success. In 2002, Eskridge took over the nuts and bolts of forming the new corporation and refining a business plan that projects annual sales of $1 billion in the United States and as high as $4 billion worldwide. He also pulled together a scientific advisory board and a board of directors now chaired by Dana M. Fowlkes, a physician and microbiologist who has successfully launched three other start-up biotechnology companies. Bode and Fischer, who have retained their university positions, serve as chief science officer and chief research officer, respectively. Eskridge said the initial $1 million in seed capital came largely from local investors equally impressed by the company's earnings potential and by the prospective benefits for eastern North Carolina. Company officials plan to build their manufacturing plant in Greenville and predict an annual payroll of $15 million within five years of start-up. As Hemocellular has gotten off the ground, federal funders have renewed their interest. The Office of Naval Research recently awarded Bode a $270,000 grant to help refine the process so it can meet FDA requirements and be scaled up to produce quantities necessary for clinical trials. Fischer, meanwhile, received funding from the National Institutes of Health for a new but related line of experiments. He's testing whether drugs used to treat hepatitis C and hemorrhagic fevers can be attached to the freeze-dried platelets. If so, it would set up a potential two-for-one treatment against some of the world's deadliest diseases. Still, significant challenges lie ahead. Eventually, Hemocellular will need at least $5 million in financing to complete the first phase of clinical trials and 10 times that to be ready to produce the first commercial vial of StasiXT. So far, fund-raising has been hampered by a slow economy. "If this deal weren't as unique as it is, we wouldn't have a chance," Eskridge said. Dr. Bode at the all-important "Freeze Mobile" freeze-dryer that he has used for the past 10 years to lyophilize StasiXT platelets. The team also must prove the technology for medical use. Freeze-dried platelets must be shown to be both safe and effective in humans. The first clinical trials, Bode said, will likely involve open-heart surgery. "Open-heart surgery patients bleed a lot," he said. "About 15 percent will bleed to the point where they're in danger of bleeding to death. We think that if you infused our products, you'd stop the blood loss and avoid shock. [Patients] will get off the table sooner and recover faster. There'll be less stress on the body." Because open-heart procedures are so carefully controlled, he said, they will provide an ideal test. The potential uses are far broader. Every year some 4 million hospital admissions involve hemorrhaging, and 50,000 people bleed to death, according to the Centers for Disease Control. That's just in the United States. Worldwide, 110,000 women will bleed to death every year in childbirth alone. Of all the potential uses, however, never far from Bode's mind is the dire need that first launched the research. "We want to get into the trauma market," he said. "Will we be able to stop death from exsanguination on the battlefield? Right now, they can only put in plasma, which will increase blood pressure and cause further bleeding. It keeps the brain alive, but you bleed out. Put in our stuff, and you save a life." That little vial of white powder begins to look pretty big. Garnet Bass is a freelance writer living in Raleigh. She is editor of edge, ECU's magazine of research and creative activity. Reprinted with permission of East, the magazine of East Carolina University, from the Winter 2004 issue, Volume 2, Number 2. 2004 Hemocellular Therapeutics, Inc. a.. 919-341-0303 b.. 919-484-0364 c.. name="platelets_final1.jpg" Content-Transfer-Encoding: base64 [ AUTOMATICALLY SKIPPING BASE64 ENCODING! ] Rate This Message: http://www.cryonet.org/cgi-bin/rate.cgi?msg=25134