Saturday, April 13, 2013

Synthesis: (S)-2-amino-3-(3-phenylacetylimidazol-4-yl)propanoic acid



Objective:
 
Synthesis of 2-Amino-3-(3-phenylacetylimidazol-4-yl)propanoic acid.
-----------------------------------------------------------------------
Materials:
Laminar flow chemical hood
Magnetic stirrer/hot plate
L-Histidine (Sigma ) 
Phenylacetyl chloride (Aldrich )
1N NaOH
Whatman #2 filter paper
dH20
Thermometer
400ml beaker
1000ml Ehrlenmeyer filter flask
Buchner funnel
Evaporation plates (2)
-----------------------------------------------------------------------
Procedure:

47.32g His
~48 ml PAC
10:00am - 11:30am
Rx in 400ml beaker w/stir bar
Temp. = 50*-65*C
7.5ml (500ul aliquots)1N NaOH added to Rx at 11:30am
50ml H2O added at 11:35
3.25ml 1N NaOH added at 11:45am
Temp. = 55*C at 11:55
Stop Rx at 12:00
Vacuum filter (#2 Whatman paper in Buchner funnel) into 1000ml Ehrlenmeyer filter flask
Pour eluant into evaporation plates (2)
Rinse flask w/ H2O- pour into evaporation plates.
Evaporate to dryness in laminar flow hood.
-----------------------------------------------------------------------
HPLC Q.C. check:
C-18 column
Mobile phase: 80 parts H2O, 20 parts CH3OH, 1 part CH3COOH
flow rate: 1ml/min
qualitative analysis performed on three (3) aliquots:
1.) Histidine control
2.) Rx beaker - after 1 hr Rx
3.) Filtered eluant (product)
Histidine control graph shows no product peaks or phenylacetic acid peak.
Rx aliquot has unreacted histidine peak, 2-amino-3-(3-phenylacetylimidazol-4-yl)propanoic acid peak, 2-amino-3-(1-phenylacetylimidazol-4-yl)propanoic acid peak, phenylacetyl linkage on primary amine of histidine (phenylacetyl - histidine) peak, phenylacetic acid peak.
Filtered eluant (product) aliquot has only product peak - 2-amino-3-(3-phenylacetylimidazol-4-yl)propanoic acid + very small (<5%) isomer peak (2-amino-3-(1-phenylacetylimidazol-4-yl)propanoic acid)
 ---------------------------------------------------------------


Thank you.

Regards,

Dennis Wright
drwright442000@yahoo.com

Thursday, April 11, 2013

Guest Article - Asbestos.com



Why the Right Mesothelioma Specialist Can Make a Difference


The most important decision a newly diagnosed mesothelioma cancer patient will make is selecting the right treatment center.
It could mean the difference between life and death.

As the experts continue unraveling the intricacies of this rare but complex cancer, novel therapies are beginning to emerge, moving mesothelioma from its gloom-and-doom past into a more-treatable future.
Advancements are being made in a variety of fronts. Diagnostics are becoming more exact. Chemotherapy drugs are getting more effective, treatments more targeted and surgeries safer.
And only a specialist will know for sure.
“This is an exciting time,” said renowned mesothelioma researcher Michele Carbone, M.D., director of the Hawaii Cancer Center. “We’re learning more how to personalize the medicine, and it’s getting better. Every case can be treated differently.”

Mesothelioma is the cancer caused almost exclusively by asbestos, often times from a long-ago exposure. An estimated 3,000 Americans each year are diagnosed.
The key for a patient and their family is finding a specialist with experience in treating mesothelioma, and a medical center with the expertise to handle it. Because mesothelioma is so rare, the majority of medical professionals rarely sees it and are ill-equipped to provide the best possible care.
Even many oncologists aren’t sure what to do. Seeing it once a year isn’t the same as seeing it every week of the year. Nothing beats experience. Only a decade ago, a diagnosis of mesothelioma came with an average survival rate of nine to 12 months.

Today it’s not unusual to see a five-year survivor. Some are living 10 years with mesothelioma. There still is no definitive cure, but the latest multimodal approach to treatment is allowing patients to live considerably longer.
Because early symptoms often mirror those of less serious illnesses, mesothelioma often is slow to be uncovered. And it can have a latency period of up to 50 years after exposure to asbestos before obvious symptoms appear.
The first critical step is getting the correct pathologic diagnosis. The next issue is staging the disease accurately, and then delivering the stage-appropriate treatment. All of it requires experts in different fields, which isn’t always found in cancer centers.

“We need to focus on getting these patients to the specialty centers,” said Joseph Friedberg, M.D., chief of thoracic surgery at the Abramson Cancer Center in Philadelphia. “It’s such a rare disease that there aren’t a lot of pockets of expertise, but that’s where the patients need to be. It can make a huge difference.”
There are centers of expertise across the country, but they are clustered in the major metropolitan areas. Centers in Boston, New York, Philadelphia, Pittsburgh, Washington D.C., Los Angeles, Dallas, Denver, Seattle and Tampa are not always easy for patients to reach. Yet that is where the expertise is, along with the latest clinical trials and novel therapies.

The patient advocates at The Mesothelioma Center can help find the specialist closest to you and the one that best fits your personalized needs.

Author Bio: Tim Povtak has been a writer with Asbestos.com since 2011.

Monday, March 18, 2013

FDA Drug Approval Process

FDA approval process

If the FDA gives the green light, the "investigative" drug will then enter three phases of clinical trials:
  • Phase 1 uses 20-80 healthy volunteers to establish a drug's safety and profile. (about 1 year) 
  • Phase 2 employs 100-300 patient volunteers to assess the drug's effectiveness. (about 2 years) 
  • Phase 3 involves 1000-3000 patients in clinics and hospitals who are monitored carefully to determine effectiveness and identify adverse reactions. (about 3 years)
The company then submits an application (usually about 100,000 pages) to the FDA for approval, a process that can take up to two and a half years. After final approval, the drug becomes available for physicians to prescribe. At this stage, the drug company will continue to report cases of adverse reactions and other clinical data to the FDA.
The research-based pharmaceutical industry currently invests some US$12.6 billion a year in new drug development. Historically, the drug development figure doubles every five years.

See also: How Drugs are Developed and Approved

Friday, June 15, 2012

Proofs


Introduction to Proofs

Proofs are the heart of mathematics. Mathematics is a prerequisite for science… and understanding science is necessary for life.
If you are a math major, then you must come to terms with proofs--you must be able to read, understand and write them. What is the secret? What magic do you need to know?
There is no secret, no mystery, no magic. All that is needed is some common sense and a basic understanding of a few trusted and easy to understand techniques.

The Structure of a Proof

The basic structure of a proof is easy: it is just a series of statements, each one being either
  • An assumption or
  • A conclusion, clearly following from an assumption or previously proved result.
 Directly and inevitably to the desired conclusion- without any distractions about irrelevant details. Each step should be clear or at least clearly justified. A good proof is easy to follow. And that is all.
Biological effects of S-2-Amino-3-(3-phenylacetylimidazol-4-yl) propanoic acid:
HEP G2 cell viability test -

DMEM
0
1
5
10
25
50
[mM]








0.215
1.311
1.275
1.16
0.864
0.247
0.187
#1
0.21
1.333
1.403
1.049
0.874
0.251
0.185
#2
0.221
1.274
1.386
1.126
0.842
0.271
0.185
#3

1.264
1.332
1.042
0.797
0.253
0.189
#4








0.2153
1.2955
1.349
1.09425
0.84425
0.2555
0.1865
average
0.005508
0.032151
0.057879
0.058048
0.034219
0.01063
0.001915
Std.Dev










Additional detail:  each well contains 200ul. HEP G2 is the ATCC 'gold standard' for a hepatocellular carcinoma (liver cancer) cell line. Concentrations are expressed in millimolars [0mM], [1mM], [5mM], [10mM], [25mM], [50mM].  Column labelled 'DMEM' is only growth media - with no cells. The test wells all have 2,500cells per well. Microtitre plate was incubated for 3 days at 37*Celsius. Color developed with MTS (a tetrazolium salt)-commonly used for cell viability determinations. The plate was read on a microtitre plate reader at a wavelength of 492nm. Resulting absorbances indicate complete (100%) inhibition of HEP G2 at a concentration of [25mM]  (S)-2-Amino-3-(3-phenylacetylimadazol-4-yl)propanoic acid.

Rapid inhibition of difficult to treat cancer cell lines.
Independently confirmed, reproducible, effective and undisputed...
 

 quod erat demonstrandum

And that is all.


Tuesday, May 22, 2012

For myself I am an optimist -

Received earlier:

 "   Hello Dr. Wright, my name is Dave i have a 7yr old friend, his name is Gage Driver. He is suffering from a tumor they call it DIPG he's  currently going to Gillett Childrens hospital in Minneapolis, with not much success. I know Chemo/Radiation along with avastin and decadron on a 7yr old isn't the answer. Please share with me what you would do, because it's only a matter of time before my friend starts that all too unpopular tailspin, that has only one ending. I've read how you claim to have a cancer stopper but why aren't people paying attention? I myself have just returned from Mexico where stemcells have lengthened my life, I suffer from a condition called alpha one anitripsin defiecency, This procedure isn't legal in the states but yet yours is, Why sir are you sitting around? My 7yr old friend wants to be president,,,now you don't want to obstruct this countries eminent future,do you. I'm hoping you can find a way to tell me one good reason you can't help us out.  Your friends Gage and Dave "

My reply: 

"Dear Dave and Gage,

Thank you. Your message is understood. I ask you to understand that we are in the process of getting this new chemical entity approved for use world-wide... and yes, there is a procedure which must be followed:

FDA approval (link updated 18 March 2013) 


I have been and continue to advance the 'state-of-the-art'.  With sufficient funding,  this new class of alpha amino acids can be in human clinical trials as soon as Summer 2012. This is all about timing and finances. Yes, many lives will be saved and many more will be spared the consequences of a cancer diagnosis.

Others are also waiting:

I heard about your research from my cousin who is a nurse.  I had no idea I would ever possibly be talking to you.  The word "on the street" is that you are using amino acids to cure cancer but that your clinical trials won't be starting until next year.   Unfortunately I don't have that long to wait.

I had a tumor on my L4 and 11 lesions on my liver as of Sept 8.  I had 10 treatments of radiation on my back and that tumor is dead.  I then had 3 courses of chemo, Gemzarine and Taxotere which did absolutely nothing for my liver, I now have numerous lesions so this is a fairly aggressive cancer.

 I am very interested if you think you can help me, I was extremely healthy until this hit.  I didn't even have any symptoms from the liver.  If it hadn't have been for the tumor on my back, I wouldn't even know there was a problem.


Yes, Dave and Gage, this does work. This compound , (S)-2-Amino-3-(3-phenylacetylimidazol-4-yl)propanoic acid, kills cancer cells - with no demonstrated toxicity to healthy cells. No surgical excision, no chemotherapy, no radiation therapy.
The only impediment to advancing the approval process is funding. I have been doing this for years - with my own personal funds and they are almost exhausted. As soon as funds are available this compound will go to GMP synthesis, CRO toxicology... and human clinical trials (Phase I - Phase I/II hybrid) can start as soon as six months afterward. This compound is eligible for expedited approval, compassionate usage exemption, accelerated review, orphan drug status. Once again, we need finances to advance the process.
http://cancercure-d.blogspot.com/



Thank you, again. Keep me apprised of events with yourself and Gage.

Sincerely,

Dennis Wright "

FYI - Gage died April 2012 from DIPG
*****************************************
Published - with permission:  
 "If I do get to the end of the next part - which I intend to be the last in that series - I won't mind if you reprint it as will be, or in some form modified in a way that's mutually agreeable. I am certainly all for treatments that don't have the toxicity of chemotherapy or the dangers of radiotherapy, but I don't want to support an approach that suggests that a cancer cure may be just round the corner. I don't believe that for a moment. Naturally anything that advances cancer treatment, palliative or curative, I could hardly object to.

 With best wishes,

Denis."
--
http://deniswright.blogspot.com/


Reality bites 5b: the sharp edge bluntly

reality 1 | reality 2 | reality 3 | reality 4 | reality 5a | reality 5b

[to finish the story....maybe.]

How does that fit in with my chess game? In chess, as the end of the game itself approaches, the losing player's king is placed in check, and the final trap is sprung when the king is checked and has nowhere to go. That's it. Checkmate.

There is one slim hope for the losing player, and that is a stalemate, but I won't try to explain that here. Well, I did explain it, but it got too complicated and didn't add anything to this story, so a pointless paragraph is gone, and we're all the better for that.

In a stalemate, no-one wins. It's a Get Out of Jail Free card for the player who was going to lose. (Ah, sorry – I've just Monopolised my chess game....)

In my game, there will be no stalemate. Mr C won't fall for it. I may evade his attack for as long as I can, but a stalemate won't happen. I know this because of the signs that are constantly increasing in number. The return of seizures, the headaches, loss of balance and increased difficulty in walking and swallowing, the strange, apparently random tremors in other parts of my body when I'm sitting or lying down; the increasing failure to remember something that was in my mind a minute ago, whether a word or an idea. Some of these symptoms of accelerated tumour activity are not completely new, but the permutations and combinations tells their own story.

It seems the king is rapidly getting boxed in.

♖     ♖     ♖     ♖     ♖

In a real game, the losing player will see defeat coming, shake hands with the opponent, and resign the game before having to play it out to an inevitable and perhaps humiliating conclusion.

Mr C doesn't like that ending. He may play a mean game of chess, but he only seeks growth at the expense of dependency, and the great irony is that his win is his own death. He will refuse to accept the resignation of his host and he will demand that the game be played out to the bitter end.

It may well be that he's more subtle than a mere biological cell-cloning program, and is capable of tiny mutations that render yesterday's treatments ineffective, or less effective than they were. His only intelligence is to find ways past the barriers that contain him and his influence. Don't be fooled; he may well be better at that game than many give him credit for, and this means researchers can be trapped in relying on outdated remedies or approaches and faith in faulty data. But that too is another story and takes me away from this one.

Here's the blunt bit. There is no honorable resignation for me. Our society, for all its multiplicity of reasons, some logical and some idiotic, decrees that the game must be played out to the last gasp. It allows no right for the player to decide just when the game should end, and thus, on grounds of higher purpose, denies the last shred of dignity in the process. And this is specially true in the sequence of events in dying from brain cancer, or other neurological calamities for the organism, where the invasion is into the core and very centre of our being.We are no longer who we were.

I've always accepted that life, by its very nature, is not fair. I go along with that. In the natural world, fairness is not an issue; for humans, fairness is a rather simplistic idea constructed by the mind, and exists only there. If you believe in fairness or unfairness in such cases, then you have the sticky question of explaining why it happened, morally – and most of the answers I've seen to that question are far from convincing. In fact, I'll go so far as to say they usually insult my intelligence.

So to me there is a terrible cruelty, with no redeeming feature, in cloaking the right to a dignified ending to the game in platitudes, specious arguments and blind dogma. None, including bishops and those new knights of the realm, our politicians, have any right to impose this nonsense upon those who do not accept their views. They play their games with our lives; but not content with that, with our deaths as well.

This didn't end up quite as you expected, did it? Me neither."

 February 2013

I don't usually make comments as I have always felt that the blog is 'Den's thing'.

I always appreciate your insightful comments Bob, and the stories you add to the mix. I fear that if other people were to contribute postings, the result would be just as much time and energy spent this end, in formatting and commenting on them - we all know he couldn't resist :-)

More importantly, the blog would no longer be about Denis and his journey, which has been very much the point.

Denis has spent a long time now, years actually, keeping everyone else informed and also entertained. I know you all appreciate how much time and effort this takes, not to mention answering the countless private emails and messages that this blog generates.

The blog has brought so many outside people in, who now feel very close to Denis and his circumstances. People that he hasn't spoken to in many years and probably may never have again, if things were 'normal'. That has been a wonderful thing for Denis and also, I feel, for them.

But then there are the people who are actually, physically here. Den's time is just as precious to us as it is to the many people who now feel that, in some small way, they own this journey as well.

So now, the selfish statement of my heart....

I sincerely hope that you would all wish, or at least understand, that as things progress into the end times that it will finally be a time for it to be just about Denis....and me....and his family.

Trace xo
************************************************************
.

For myself I am an optimist - it does not seem to be much use being anything else.
Sir Winston Churchill

Tuesday, April 24, 2012

Guest Article - The Mesothelioma Center at Asbestos.com

 






Breakthroughs for Mesothelioma Cancer Research
For patients with mesothelioma, treatment options are often limited. Traditional surgical treatments such as extrapleural pneumonectomy (EPP) and pleurectomy/decortation (P/D) are radical surgeries that may only provide a few more months of survival to a patient.
Surgery is often combined with other treatments such as chemotherapy and radiation, but mesothelioma is notoriously resistant to chemotherapy. Scientists are researching new methods of treatment on the molecular level in the form of mesothelioma biomarkers, suicide gene therapy and personalized treatment through DNA scanning.
   
Biomarkers
Biomarker research has been a promising area of advancement in the pursuit of new treatments for mesothelioma. A significant recent development has been the discovery of mesothelin as a mesothelioma biomarker. It is a naturally occurring glycol-protein found in normal mesothelial cells. In mesothelioma patients, it is overexpressed by mutated cells.
The results of a recent Phase I clinical trial revealed that a mouse-human chimeric monoclonal antibody called MORAb-009 could create cytotoxicity and kill mesothelin-expressing cell lines. Meaning this antibody could kill malignant mesothelioma cells.
   
Suicide Gene Therapy
Mesothelioma, like other cancers, resists apoptosis. Through gene therapy, researchers have begun to induce apoptosis in cancer cells through suicide gene therapy.
Because mesothelioma is a localized tumor in early stages, rapid and efficient delivery of genes could be accomplished through the layer of mesothelial cells. The herpes simplex virus-1 thymidine kinase (HSVtk) gene is often used to induce tumor cell death.
In a study, the virus gene was administered to 30 patients with minimal side effects. A number of patients survived for more than three years, including a patient who was still alive after 10 years.
   
DNA Scanning
Because a mesothelioma patient’s prognosis is tied to many individual factors, a single solution does not often help every patient. Foundation Medicine, Inc. has pioneered a new high-speed DNA scan based on comprehensive cancer genomic testing. 
Foundation’s high speed DNA scan has isolated 200 genes with a known link to cancer. The test can match cell abnormalities from a specific tumor to a drug that can attack the abnormality.
Using this new technology, scientists isolated a genetic flaw in 2 percent of 561 lung cancer tumors. It was matched with a kidney-cancer drug named Sutent and a thyroid-cancer drug called Caprelsa. Scientists found that mutated cancer cells treated with these specific drugs died. This type of targeted therapy has positive implications for mesothelioma patients.
As researchers continue to test the boundaries of traditional cancer therapy, mesothelioma patients can be hopeful for more effective therapies.
   
Bio: Michelle Y. Llamas is a writer for the Mesothelioma Center. She is committed to generating mesothelioma awareness and providing information regarding breakthroughs in mesothelioma treatment.

   
Sources:
Hassan, R., Schweizer, C., Lu, K. F., Schuler, B., Remaley, A. T., Weil, S. C., & Pastan, I. (2010). Inhibition of mesothelin-CA-125 interaction in patients with mesothelioma by the anti-mesothelin monoclonal antibody MORAb-009: Implications for cancer therapy. Retrieved from http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2864325/?tool=pubmed
MIT. (2011). Foundation Medicine: Personalizing cancer drugs. Retrieved from http://m.technologyreview.com/biomedicine/39707/
Tannapfel, A. (Ed.). (2011). Malignant mesothelioma: Recent results in cancer research. New York: Springer.
Vachani, A., Moon, E., Wakeam, E., & Albelda, S. M., (2010). Gene therapy for mesothelioma and lung cancer. Retrieved from http://www.ncbi.nlm.nih.gov/pubmed/20160042