cryonics continued...

john grigg (starman125@hotmail.com)
Wed, 29 Sep 1999 15:30:11 PDT

Hello again,

And now for a post by Mike Darwin and one by Robert Ettinger.

Date: Tue, 28 Sep 1999 01:19:37 -0400
From: Mike Darwin <75120.575@compuserve.com> Subject: 21CM Patent: Some answers and questions

There have been a number of comments and speculations on the recently published 21st Century Medicine (21CM) assigned patent on novel cryoprotectants. In particular, Robert Ettinger and Thomas Donaldson have expressed puzzlement about the lack of supporting data for the high rates of cooling claimed, and also as to the "legitimacy" of the core patent claims themselves. For instance, Ettinger writes (as quoted in the full text of his message below): "The inventors acknowledge that glycol ethers have been used before to preserve embryos and cell suspensions, but believe they have priority in application to tissues, organs, and animals. Obviously, there are potentially interesting questions in patent law--both as to the relation between these inventors and prior users of glycol ethers, and between these inventors and later users of new variations and applications." Donaldson also states: "There is also some question in my own mind about whether or not some of the perfusion methods described will prove to be patentable ..."

Ettinger also notes that he is "legally untutorted" (sic) in patent law.

This last observation is a critical one. *Claims* that at first blush seem broad and overweening may on careful scrutiny be narrower than someone inexperienced in reading (and circumventing!) patents may percieve. The patent issused to 21CM and created by Wowk, Darwin, Russell and Harris acknowledges prior art in the use of alkyoxylated organic molecules as cryoprotectants as they both morally and legally should and must (willful failure to acnowledge prior art is grounds for voiding of a patent). The seeming "iota" here upon which the claims rest is to *vitrification* as opposed to freezing. And that is no iota at all.

The cryonics community as a whole has long been confused about the difference between these two approaches to cryopreservation, and I do not intend to spend time in further, and probably futile, attempts at their education. Suffice it to say that ice forms crystals which greatly and perhaps irreversibly damage (destroy information) in tissue, Merkle and Tipler notwithstanding ;-). Alkyoxylated cryoprotectants still confer advantages in freezing where ice is formed over conventional cryoprotectants in many situations.

However, that is *not* the application that is being claimed in the 21CM patent. Rather, it is the use of these compounds in clever ways to achieve *vitrification*: cryopreservation in the absence of damaging amounts of ice, wherein the organ system or tissueis maintained in a largely or completely glassy or otherwise non-crystalline state. Freezing is a workable method for cryopreservation of of many cells, small organisms, and tissues, but fails miserably when applied to complex, interconnected masses of cells ans tissues (such as mammals and most of their their major solid organs).

Additional confusion results from the mechanics of patent law and the patent process which requires *multiple* experiences with in order to even begin to understand how it operates. In other words, you need to have dealt with *good* patent attornies and prosecuted *multiple* patents before you get a feel for the quirks of the system. FYI, as an aside, this is a process that will cost you many, many thousands of dollars. It is often said that a physician who has himself for a patient is a fool. The same can be said for those who step into the byzantine world of *serious* patent prosecution without the best counsel available.

One thing that is commonly done is to (seemingly rationally enough) link integerally related novel technologies together in a single patent. The U.S. Patent Office often has other ideas and will break a single patent into several parts, or into several discrete patents, each of which will issue (and finally post to the Federal website) at different times. (There are reasons for this: each patent generates Federal revenue and must be maintained by regular payments over its life). Thus, a large section of the 21CM patent currently under discussion on Cryonet has not yet posted to the web. This section describes technology developed largely by Darwin and Wowk to achieve very high rates of cooling in whole mammals via intravascular perfusion with a variety of "inert liquids." Brian Wowk deserves particular credit for discovering some unique and truly unexpected properties of perflurochemicals which allow continued non-toxic perfusion (rapid intravasclar circulation) of these agents at temperatures as low as -150 C. This is important, because it allows not only for very fast rates of cooling, but is below the glass transition point (Tg) of most water-cryoprotectant mixtures. This, in effect, eliminates the danger of ice formation during cooling and rewarming due to "devitrification." We expect this patent to post later this year.

I will comment specifically on the full-text of Ettinger's post, and on a relevant sentence in Dolaldson's post:

>Message #12443
>From: Ettinger@aol.com
>Date: Mon, 20 Sep 1999 14:08:22 EDT
>Subject: 21CM patent

>Go to www.uspto.gov to access the full text of the patent (Sep.14)
assigned
>to 21st Century Medicine Inc. Oddly enough, the inventors listed do not
>include Greg Fahy; they are Brian G. Wowk, Michael G. Federowicz
["Darwin"],
>Sandra R. Russell, and Steven B. Harris. It runs about 18 printed pages.

There is nothing odd about the omission of Greg Fahy from this patent. The work which this patent was based upon occurred as a result of a truly bizzare chain of circumstances which began with a request I posted to Cryonet well over two years ago. I explained in that post that BioPresevation (my company, which at that time was active in postmortem human cryopreservvaion) had a client who's brain had been fixed in formlain and been refrigerated, but not frozen, following her legal death. There was no possibility of achieving vascular access and introducing cryoprotectants.

I asked for advice from all quarters on what might done in such a situation, and in particular if anyone had data on more highly permeable cryoprotectants than those in conventional use (primarily glycerol and DMSO). Only one person responded to my request for help and that was Doug Skrecky. Doug played a critical role in all that was to come by providing me with a list a of a few highly permeable likely candidates for cryoprotection, *and* a reference paper which listed the red cell permeabilities of well over a hundred organic compounds. This list was daunting, but exploration was begun, initially using a mixture of physical chemistry assays (concentration needed to vitrify, or CNV) and a simple and inexpensive toxcological assay I developed while in high school.

Meanwhile, I had forwarded this paper to Brian Wowk who at that time was still working on his dssertation in Medical Physics in Winnipeg, Canada. Brian selected a number of high permeability compounds to test which he felt, based on their stucture (and his insigtful understading of water-cryoprotectant physical chemistry) might be good candidates. One of these candidates, 2-methoxyethanol, behaved in a fashion unlike any cryoprotectant I had observed before. It vitrified at anamolously low concentrations, it penetrated the entire brain rapidly and uniformly, including the myleinated axons which have been a major barrier to successful brain cryoprservation, and it passed my bioassay tests, including human RBC freezig and vitrification studies which Sandra Russell and I ardously carried out over a period of several months. This molecule, and related ones, also exhibited gross physical behavior unlike anything I had observed in any class of chemicals which had been tested as putative cryoprotectants. Space prevents me from describing these phenomena, but they electrified both me and Steve Harris.

Brian then took up the torch in Winnipeg in the midst of his dissertation and began systematically evaluating mathoxylated compounds in the basement of his home using equipment I had sent him and a test tube from his childhood chemistry set. Brian was later to elegantly discover using a $50K differential scanning calorimeter, instread of a single test tube from an ancient chmeistry set, that each methoxyl group added to most suitable cryoprotectant molecules decreases the CNV by a few per cent. This is no small matter, since a difference in concentration of 1% cryoprotectant can mean the difference between near complete loss of viability (and serious ultrastructural disruption) and virtually 100% survival for organs such as the mammalian kidney. The rapid penetration (equilibration) of these agents and their greater depression of the freezing point offered additional opprtunities to capitalize on the decreased CNV requirement: shorter loading and unloading times at lower temperatures were in theory possible, with resultant further reduction in toxicity, or CNV, or both.

During this time Greg Fahy was working for a competing company and IMHO thought little of our ability or efforts to make significant progress at 21CM. In fact, he was working with a company with whom 21CM had anything but cordial relations. BOTH sides dealt with each at *more* than arms length, and the nandwidth of communication was zero.

Greg was not appraised of the new approaches being developed by 21CM for nearly two years -- and considerably after the first disclosure document for the patent had been filed. Greg can, of course, speak for himself, but I think he was quite astounded at the progress that had been made completely independent of his efforts or inputs (aside from those, such as the basic principles of vitrification, which had been openly published by Greg in the literature).

I want to pause here in this narrative and thank Doug Skrecky publically for his generosity and his intellectual insight in suggesting that we consult the critical reference that lead on to much of what has happened since. I also want to make very clear that the intellecual property in this patent was the work of three people: Wowk, Darwin and Russell. Each made pivotal contributions and spent countless back-breaking hours in the laboratory.

Ettinger continues:

>Naturally, we will investigate the procedures and substances mentioned.

>There are many interesting features in the patent, not least the structure
of
>claims. The claims start broadly, including any alkoxylated organic
compound
>in a concentration sufficient to permit vitrification and cooling until
>vitrified; and the application of this to any organ, tissue, or animal.
More
>specific claims relate to use of 2-methoxyethanol and
>3-methoxy-1,2-propanediol.

>The inventors acknowledge that glycol ethers have been used before to
>preserve embryos and cell suspensions, but believe they have priority in
>application to tissues, organs, and animals. Obviously, there are
potentially
>interesting questions in patent law--both as to the relation between these

>inventors and prior users of glycol ethers, and between these inventors
and
>later users of new variations and applications.

"Summary" of the invention mentions perfusing an inert fluid through the vascular system and controlling the temperature of the fluid; and a class of
new glycol ether CPAs. Oddly enough, the Claims do not (to my legally untutored mind) appear to make any direct or specific reference to cooling methods, even though in the "Summary" that seems important.

>"Glycol ethers" are understood to include compounds containing alkoxy--and

>particularly methoxy--functional groups. Included are alkoxylated alkanes
and
>alkoxylated alcohols and polyols, with several sub-groups given.

>For some organs, cooling and rewarming rates can exceed 100 degC per
minute.
>[This seems to suggest possible use with other CPAs, such as amides.]

>"Large animals can be perfused wit high concentrations of glycol ethers
near
>0 C with rapid equilibration, no dehydration, no edema or other visible
>evidence of toxic effects. Histologic preservation is excellent at
>microscopic and ultrasturctural levels."

>References include Skrecky's proposal in Cryonet #5174, 1995; CRYONICS:
>REACHING FOR TOMORROW (Wowk & Darwin, 1991; and THE PROSPECT OF
IMMORTALITY,
>1964; and a Darwin/Hixon piece in CRYONICS July 1984, as well as various
>patents and publications in recognized journals.

>I have not found any patent entries under "ice blockers," although the
>instant patent says that suppression of ice formation is an important
>feature. I believe 21CM has a different class of substances it calls ice
>blockers, acting through a different mechanism.

>The "Conclusion" mentions unprecedentedly high rates of cooling and
>rewarming,;and the advantages of the new CPAs, including penetration and
low
>viscosity, rapid equilibration, ice inhibition, minimization of toxicity,
and
>glass forming properties.

>Congratulations to all the inventors involved. Lots of work ahead.

>Robert Ettinger
>Cryonics Institute
>Immortalist Society
>http://www.cryonics.org

What also needs to be mentioned is that during tail-end of this same period of time a South African (SA) woman named Olga Visser appeared on the international scene making exatrodinary claims for a novel highly penetrating and nontoxic cryoprotectant which reputedly allowed for successful liquid nitrogen temperature (-196 C) cryopreservation of rat hearts. Unbeknownst to most American cryonicists, Ms. Visser was also claiming that a similar (and it turns out identical compound) was a cure for AIDS (HIV infection). This actually is what brough Ms. Visser to my focused attention. I count as friends and colleagues a number of critical care physicians ("Intensivists") in South Africa. Several of them were distressed beyond words at the actions of Ms. Visser and her husband Ziggy Visser in facilitating the replacement of the respected (and highly flexible and decent) chair of the South African Medicines Council (the approximate equivalent of our U.S. FDA) with a woman with less medical understanding than a village Medicine Man. Using political entre and the Medicines Council's new chief, the Visser's managed to route "Virodene" around all normal channels of peer review, clinical trials, and informed consent. Virodene was, my SA colleagues alleged, being given to human HIV patiets in violation of both the Nuremberg and Helsinki codes.

I was asked for information on the matter because her connection to the American cryonics community and her claims regarding rat heart preservation were being bandied about in the SA media at the time. The Intensivists who knew me and of my involvement in cryonics and cryobiology asked for my opinion as to her credibility. They also, without my asking, provided me with the name of the compound Ms. Visser was using as her cryoprtectant and AIDS cure: dimethylformamide (DMF). This information was obtained, as I understand it, from a purchasing agent at the hospital in the perfusion department where Ms. Visser was employed.

DMF was one of the many molecules on the list Doug Skrecky had lead us onto. We had examined it and found it be both toxic in cryoprotective concentrations, but far more to the point, very poorly effective as a cryoprotectant in freezing, and a complete write-off for vitrification. In fact, DMF caused the formation of some of the largest ice crystals and the most severe organ damage on freezing of *any* of the conventional cryoprotectants we had evaluated it in comparison to. We had, in short, discarded DMF as a viable highly permeating mono-agent for organ or tissue cryopreservation. We had also investigated many related compounds (peralkylamides)and also found them to be of no utility in our models. I communicated this to my SA colleagues and to the Reuters and other reporters who subsequently called. Similarly Charles Platt was instrumental in informing key members of the international press of Visser's likely credibility and, most importantly, of the identity of the industrial solvent, immunosuppresive, and carcinogen (DMF) she was promoting as an AIDS cure

Ms. Visser was ultimately discreited and left SA. However, her legacy remains: human clinical trials, even of the most ethical and tightly controlled nature, have become all but impossible due to the "Virodene Scandal" or the "Visser Affair." My colleague and friend Dick Burrows, Diector of Intensive Care Medicine at Addington Hospital in SA, to this day remains bitter and inflamed about the "enormous harm" the Virodene fiasco did to clinical and experimental medicine in SA. He has repeatedly stated that his practice of medicine has suffered direct harm as a result of this incident. (Dr R Burrows, Director Intensive Care Unit, Addington Hospital PO Box 977 PO Box 977, Email burrows@csurf.co.za).

I might also note that money and support from cryonics organizations in the US such as Alcor and CI ( I've been told in the vicinity of $50,000 to $75,000) helped to allow Visser to continue her fraud in a country where the level of desperation for ANY relief from AIDS simply cannot be understood by us in the US. FYI: roughly half of all the babies born in Dr. Burrows' hospital are HIV infected.

In fact, 21CM recently abandoned efforts at clinical trials for promising drugs to treat shock and multisystem organ failure from trauma and sepis which were ideally suited to a "Third World " environment and economy *directly* as a result of the Visser affair.

Ettinger also notes:

">I believe 21CM has a different class of substances it calls ice
>blockers, acting through a different mechanism.

This is correct. This work, begun by Brian Wowk, has continued to progress at an astounding pace. In no small part this been due to truly hurculean efforts by Eugene Leitl, who I have seen putting in day after, day after day of 14, 16 and even 20 hours at a stretch working to synthesize novel ice blocking molecules: a task in which he has succeeded remarkably!

Thus, the patents which are "posting" now (i.e., becoming web accessible) are but the tip of the iceberg, or should I more appropriately say, "the first vitreous prearl of a mountain of precious, ice free glass which is yet to emerge."

Thomas Donaldson comments:

>I too look forward to wider application of the cryoprotectants and
>perfusion technology created by 21st Century Medicine. Yes, it may not be
>immediate; the first thing to do is to try these methods on brains.
>(There was a time lapse between the first Wright brothers flight and
>the use of 747's, after all).

This has been done, and with stellar results and has already been reported on. In fact, ACS has pictures on their website.

As to cooling, Thomas was apprised of the fact we had achieve cooling rates of 10 C/min to -70 C with intact 20+ kilo dogs at the A4M meeting in a private brefing held *at least* three years ago, if not longer.

Furthermore, the financial and technological barriers to the application of this technology to an estimated 1/3rd of human cryonics patients NOW, are trivial compared, say, to the effort that has gone into the construction of CI's latest and largest cryostat or Alcor's ambitious program of training and "suspension services" revamping.

And the ultimate irony is, the storage costs will probably be lower and the up-front technology not more than 20% to 30% more costly than that employed by Alcor now (the only high technology player left in the human cryopreservation field).

As a result of all this, I have become convinced that the only thing that will get the impact of the progress *already made* through the thick skulls of cryonicists is when they hear about the revival and cure of the first *human* cancer patient placed into suspended animation at -120 C or below.

I predict the response will be: "Don't waste your money on it, get *frozen* on the cheap and wait till you can get reanimated for free and be fantastically rich and sexually irresistiable using Nanotechnology."

Mike Darwin, Director of Research
Critical Care Research, Inc.
10743 Civic Center Drive
Rancho Cucamonga, CA 91730
Phone: (909) 987-3883
Fax: (909) 987-7253
email: 75120.575@compuserve.com

PS: My thanks to John Grigg for his warm remarks:

>I looked over the patent for 21st Century Medicine's research developments

>and must say I am VERY IMPRESSED!!! I congratulate those who made it
>possible!!

From: Ettinger@aol.com
Date: Tue, 28 Sep 1999 11:10:37 EDT
Subject: 21CM, ABS etc.

Mike Darwin's post today (CryoNet #12464) included the following:



Ettinger also notes:

">I believe 21CM has a different class of substances it calls ice blockers, acting through a different mechanism.

This is correct. This work, begun by Brian Wowk, has continued to progress at
an astounding pace. In no small part this been due to truly hurculean efforts
by Eugene Leitl, who I have seen putting in day after, day after day of 14, 16 and even 20 hours at a stretch working to synthesize novel ice blocking molecules: a task in which he has succeeded remarkably!

Thus, the patents which are "posting" now (i.e., becoming web accessible) are
but the tip of the iceberg, or should I more appropriately say, "the first vitreous prearl of a mountain of precious, ice free glass which is yet to emerge."

Thomas Donaldson comments:

>I too look forward to wider application of the cryoprotectants and
>perfusion
>technology created by 21st Century Medicine. Yes, it may not be immediate;
the >first thing to do is to try these methods on brains. (There was a time lapse between >the first Wright brothers flight and the use of 747's, after all).

This has been done, and with stellar results and has already been reported on. In fact, ACS has pictures on their website.

As to cooling, Thomas was apprised of the fact we had achieve cooling rates of 10 C/min to -70 C with intact 20+ kilo dogs at the A4M meeting in a private brefing held *at least* three years ago, if not longer.

Furthermore, the financial and technological barriers to the application of this technology to an estimated 1/3rd of human cryonics patients NOW, are trivial compared, say, to the effort that has gone into the construction of CI's latest and largest cryostat or Alcor's ambitious program of training and
"suspension services" revamping.

And the ultimate irony is, the storage costs will probably be lower and the up-front technology not more than 20% to 30% more costly than that employed by Alcor now (the only high technology player left in the human cryopreservation field).

[End of Darwin excerpt]



Darwin's last two paragraphs above (together with further comments here omitted) seem to constitute a reproach against CI and Alcor for failing to make 21CM's advances available to our members. However, not only is a great deal of information still unavailable to us, as Darwin himself emphasizes, but one of Saul Kent's new companies, Advanced BioSciences (ABS) has been formed for the specific purpose of marketing 21CM technologies and interfacing with cryonics organizations. The initiative necessarily remains with 21CM, ABS et al.

So the situation remains as I have previously described it:

First, CI will continue, within our constraints, to do our own research and to modify our procedures as may seem justified from time to time.

Second, CI will aim over time to make all options available to our members, if necessary at a higher cost for more expensive procedures (and perhaps at a
lower cost for less expensive procedures, e.g. if some combination of fixation, drying, and freezing should prove sufficiently effective in preserving structure).

Third, we will of course pay careful attention to any proposals from ABS, BT,
or anyone else with something specific to offer.

Robert Ettinger
Cryonics Institute
Immortalist Society
http://www.cryonics.org



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