人々の “健康促進” のために!

人々の “健康促進” のために!
2015年春、沖縄の琉球大学キャンパス内 (産学共同研究棟) に立ち上げた “PAK研究センター” の発足メンバー(左から4人目が、所長の多和田真吉名誉教授)
For detail, click the above image.

2009年12月27日日曜日

Edwin Krebs (1918-2009), A Great Pioneer in Kinase Research

Those who have ever studied the reversible phosphorylation of proteins would
owe much both Edwin Krebs and his colleague Edmond Fischer at University
of Washington in Seattle. In 1950s they found the first protein kinase,
phosphorylase kinase (PK), which phosphorylates and activates the enzyme
glycogen phosphorylase (GP). This phosphorylase catalyzes the hydrolysis
of glycogen, supplying the glucose essential for a variety of our actions
such as muscle contraction and learning. PK also requires the phosphorylation
by another kinase called PKA (cyclic AMP-dependent kinase). cyclic AMP (cAMP)
is a second messenger that is produced by adenylate cyclase (AC) from ATP.
AC is activated by a GTPase (or G protein) called Gs. Thus,
A G protein activates this kinase cascade through the second messenger cAMP.

A similar G protein-kinase cascade was found around 1994, when Ed Manser
in Singapore cloned the first mammalian member of PAK family (PAK1) in brain.
This kinase (PAK1) is directly activated by another G protein family (RAC
and CDC42).
Later it was found that RAC/CDC42 is down stream of another kinase (PI-3
kinase) which is directly activated by the oncogenic G protein RAS. In
other words PAK1
mediates the oncogenic signal of RAS, and is essential for the growth of
more than 70% of all human cancers including breast and prostate cancers
and NF (neurofibromatosis) tumors. In addition, PAK1 is essential for both
metastasis and angiogenesis of these solid tumors. Since PAK1 is not essential
for the normal cell growth, anti-PAK1 drugs such as Bio 30 (NZ propolis
extract) would be powerful therapeutics for the treatment of these PAK1-dependent
cancers and NF without any side effect. Thus, cancer and NF patients should
greatly appreciate the discovery of the first kinase cascade (PKA-PK-GP)
during 1950s-1960s, which eventually led to the discovery of the oncogenic
RAS-PI-3 kinase-RAC/CDC42-PAK1 signal cascade in 1990s.

Accordingly, the 1992 Nobel prize in Physiology/Medicine was awarded to
Edwin Krebs and Edmond Fischer.

2009年12月24日木曜日

RNA Editing Genes Determine
the Longevity of Human and Nematode

The (tumor suppressing) transcription factor "FOXO" is known to play the
key role in determining the life span of human and nematode. The kinase
cascade including LKB1 and AMPK activates FOXO, leading to the expansion
of life span, whereas the kinases AKT and PAK1 inactivate FOXO, leading
to the shortening of life span. There are several natural products that
both activate LKB1/AMPK and inactivate PAK1, leading to the expansion of
life span: CAPE (caffeic acid phenethyl ester) from Propolis, Curcumin
from Indian Curry, Capsaicin from Chili Pepper, Resveratrol from Red Grapes,
and Berberine. Thus, these products are among natural "Elixirs".

Recently, Paola Sebastiani's group at Boston University found that RNA editing
genes, ADR-1 and ADR-2, as well as RNA interfering gene RDE-1, also play
the key role in determining the life span of both human beings and nematodes
such as C. elegans. Dysfunction of RNA editing genes increases the RNA interfering activity of RDEs, and shortens the life span of nematode by 50%. Interestingly, however, dysfunction of RDE-1 gene could rescue the above (short-lived) nematode from the premature death. Thus, like the oncogenic kinases PAK1 and AKT, RDE-1 appears to shorten the life span by its RNA interfering activity (blocking the translation of its specific target genes which still remain to be identified).

Since Hsp16 (encoding a heat shock protein) is among the key genes downstream
of FOXO contributing to the expansion of life span, it is possible that
RDE-1 might block the translation of Hsp16 (or FOXO) RNA. We recently found that PAK1 blocks the expression of Hsp16 gene, and CAPE can re-activate Hsp16 gene
by inactivating PAK1. Also it would be of great interest to see if these natural elixirs activate ADRs or inactivate RDEs somehow.

2009年12月6日日曜日

たんぽぽの葉は、 食用 にも薬用にもなる!

http://www.geocities.co.jp/NatureLand/1351/recipe.htm

初夏になり、引っ越したばかりの我が新居の庭一面に生え始めているのに気づき、
開花前の柔かい葉を選んでは採集し、水で簡単に洗ったのち、生でサラダ代わりに
食べたり、ラーメンに入れて「山菜ラーメン」を作ったりしている。
多少苦味はあるが、野菜不足の解消に十分なるわいとすっかり満足していたら、
驚くなかれ(良薬は口に苦し!)、最近(2008年)の英文医学雑誌を調べて
みると、 西洋たんぽぽ (Taraxacum officinale) の葉 (水エキス)には乳癌
などのPAK依存性の癌の増殖や転移を抑える作用があることが、(ナバホ族の子孫が
多く住む)米国ニューメキシコ州の首都(アルバカーキー)にある大学の研究
グループによって、発見されていることが判明した。 更に10年ほど前には、
京都薬科大学の研究グループが、日本産のたんぽぽ(根)にも制癌作用がある
ことを薬学雑誌に発表している。けだし、中国では漢方薬の1つ「蒲公英」として、
昔から使用されているそうである。アラビア人も北米のインディアンも伝承薬として、
使っているようだ。 先人の英知に痛く感服した。

癌の予防にもなるが、末期の肺癌にも効く!
http://www.healingcancernaturally.com/dandelion-taraxacum-root-cures.html

Towards the Development of a New "Red Propolis"
Based on Sichuan Pepper (Hua Jiao)

A few years ago we found that Hua Jiao fruits (Chinese red peppercorns)
contain an anti-PAK ingredient which can be extracted by 70% ethanol (or
warm-water), and suppress the growth of cancers and NF tumor xenografts
in mice (Hirokawa, Y. et al, 2006). Since this extract has a rather bitter
(not spicy) taste, we tried to commercialize the dried power of this extract
blended with an NZ honey to add a sweet taste under the brand name "Honey
Pepper" as the first effective therapeutic for NF (neurofibromatosis) and
PAK-dependent cancers such as pancreatic and colon cancers, for which no
effective therapeutic was available on the market. However, no company
got interested in this project. Perhaps it might be too "inexpensive" for
the commercialization.

A year later we found that one of NZ propolis extracts called "Bio 30" also
suppresses the growth of PAK-dependent cancers and NF tumor xenografts in
mice (Demestre, M. et al, 2009). Bio 30 is an inexpensive CAPE (caffeic
acid phenethyl ester)-rich water-miscible extract of NZ propolis from Manuka
Health based in Auckland, NZ, and selectively blocks the oncogenic kinase
PAK by down-regulating the GTPase RAC. One minor problem with this extract
is that its major anti-cancer ingredient (CAPE) causes a skin allergy to
1-2% of population including myself (!).

Shortly thereafter we found that another propolis extract called GPE, Brazilian
green propolis extract from Yamada Bee Farm in Japan, also suppresses the
growth of NF tumors and PAK-dependent cancers by blocking the kinase PAK
(Messerli, S. et al, 2009). GPE contains no CAPE, but another anti-cancer
polyphenol called ARC (artepillin C) that inactivates PAK. This extract
does not cause any allergic reaction, but is far more expensive than Bio
30 (which costs only a dollar for daily treatment).

Interestingly, Brazilian "red propolis" extract (RPE) also appears to inactivate
the kinase PAK, as it suppresses the growth of pancreatic cancer, although
it contains neither CAPE nor ARC. These findings strongly suggest that
honey bees have a natural instinct to collect anti-PAK ingredients from
a variety of trees (or shrubs) such as poplar to make propolis (bee hive)
for protecting their larva from a variety of harmful pathogens such as bacteria,
fungi, viruses, etc. However, RPE is as expensive as GPE. More recently,
I heard that there is another "red propolis" in China which appears to be
based on CAPE. However, this Chinese red propolis is not available on the
market as yet.

Thus, I got a new idea that honey bees could collect the anti-PAK ingredient
from red fruits (peppercorns) of the Chinese "Hua Jiao" to produce an inexpensive
"red propolis" which would be useful for the therapy of both NF and cancers.
The key for this future project would be how to condition honey bees towards
this red peppercorn. Honey bees can memorize colors (in particular yellow
and violet), sweet taste of sugars and aromatic scent. So if the aromatic
alcohol extract of Hua Jiao is mixed with honey and spotted on a red paper,
in theory honey bees would approach this sweet aromatic red paper, learning
that the aromatic red fruits could be among their potentially favorable sources
of sweet honey. Once they could locate Hua Jiao, they would become addicted
to its anti-PAK ingredient whose chemical nature nobody has identified as
yet. Once the new "red propolis" is produced in my garden (bee farm), we
shall make the ethanol extract of this propolis to confirm its anti-PAK
and anti-cancer potential in vitro and in vivo.

However, commercialization of this OZ red propolis extract called "HJ propolis"
would eventually need the chemical identification of this anti-PAK ingredient
in this propolis (or Hua Jiao) to standardize its content in this new propolis
product. Thus, it might be a life-long (certainly time-consuming) project
towards the commercialization. Generally speaking, natural anti-PAK products
including "Natto" and propolis would have a potentially huge market value,
simply because they would be useful not only for cancers and NF, but also
several other PAK-dependent formidable diseases such as Alzheimer's (AD),
Huntington's (HD), AIDS, inflammatory diseases (Asthma and arthritis), epilepsy,
type-2 diabetes, hypertension, HCM (hypertrophic cardiomyopathy ) and malaria.

We prefer to developing an inexpensive one if possible, because it would
be far more "democratic", as it could offer a benefit not only the very
rich people, but also the very poor, even for a life-long treatment if
necessary. These diseases never discriminate the rich and the poor. So the
therapeutics should not. Thus, it would be worth trying to develop this
inexpensive red propolis in my own garden.

2009年12月2日水曜日

Civil War in Afghanistan Should be Solved
by Afghan People, and NOT by Outsiders.

Like Vietnam Civil War, the ongoing civil war in Afghanistan could not be
settled down by outsiders such as US and NATO forces or Russian/Chinese
forces. It should be ended by Afghan people, including both Karzai government
and Taliban forces.

So I strongly oppose the Obama's US government's recent proposal to send
more US troops of 30,000 soldiers to Afghanistan. It would be just the huge
waste of both young soldiers' lives and money of all US tax payers for nothing!
Besides, unlike Iraq there is no oil in this area. Only opium is available
there. His US government has many other important jobs to be urgently solved
inside the United States such as huge financial crisis, healthcare problem,
the mysterious CCD (colony collapse disorder) of honey bees, and global
warming which would definitely affect the rest of this world.

Also I would like to urge his US government to move their US military bases
from Okinawa Island in Japan to Guam Island which is within US territory.
Japanese people don't need US troops any more, which kept occupying Japanese
islands, in particular Okinawa, since the end of WWII for more than six
decades. Japan is an independent country with high-technology and strong
economic power, and Japanese people would defend their own country by themselves
if necessary (in case some other aggressive country invades Japanese soil
in the future). Interestingly, Japanese soil has never been invaded by any
other country for more than seven centuries since the Mongolian/ Chinese
dynasty tried to invade Japan in late 13th century, but unsuccessfully. It
is very unlikely that either Russian, Chinese or Korean would invade such
a tiny territory of Japan in the future, which has no valuable natural resources
except for over-crowed population (more than 120 million people).

2009年11月18日水曜日

The Most Urgent Project in Colon Cancer Research:
In vivo Proof of Anti-PAK1 Drugs' Effectiveness

Colon cancer is the "number 1" killer in Australia. The 1960 Nobel laureate
MacFarlane Burnet (1899-1985) in Melbourne, a great spiritual mentor of
mine during my university days in Tokyo, who inspired me to study the molecular biology
of immunological self-recognition, was among the numerous victims of colon
cancer. However, so far no effective therapeutic has been developed for
this formidable cancer as yet.

In 1989, a few years after his death, it was revealed that 50% of human colon cancers
carry the oncogenic mutant of RAS (mostly K-Ras), suggesting that abnormal activation
of this RAS is among the major causes of this cancer. Since RAS activates
the kinase PAK1, and RAS-induced malignant transformation requires this kinase,
it would be clear that at least 50% of colon cancers,
namely "RAS" colon cancer, could be treated effectively by anti-PAK1 drugs
such as Bio 30 which are available on the market inexpensively.

How about the remaining "non-RAS" colon cancers which carry no RAS mutant?
Around 1991 it was reported that more than 70% of human colon cancers carry
a loss-of -function mutation (dysfunction) of the tumor suppressor p53,
and the oncogenic RAS mutation and the dysfunction of p53 co-operate for
the malignant transformation, suggesting that p53 normally blocks the RAS-induced
malignant transformation. In other words, at least 70% of human colon cancers
including all RAS and 40% of the remaining non-RAS (=p53-deficient) colon
cancers could be treated effectively by anti-PAK1 drugs.

How about the remaining 30% of human colon cancers which carry neither RAS
nor p53 mutant? Around 1996 it was found that more than 90% of human colon
cancers carry a loss-of-function mutation of the tumor suppressor APC, and
a combination of both the oncogenic RAS mutation and the dysfunction of APC
is essential for the malignant transformation (either RAS or APC mutation
alone is insufficient to cause this cancer). In other words, APC normally
blocks the RAS-induced malignant transformation, and therefore at least 90%
of human colon cancers including all RAS and 80% of the remaining non-RAS
(=APC-deficient) colon cancers could be treated effectively by anti-PAK1 drugs.

However, so far nobody has experimentally confirmed in vivo that one of
the anti-PAK1 drugs available on the market can indeed suppress the growth
of human colon cancer, xenografts in mice or clinically. In my opinion,
that would be the most urgent project to be performed for identifying the
first effective therapeutics for colon cancers.

In 2003 Shigetoshi Kadota's group at Toyama Medical and Pharmaceutical University
in Japan reported that CAPE (caffeic acid phenethyl ester) and its analogues block
the lung metastasis of colon cancer cell line (26-L5) in mice. CAPE inactivates
the kinase PAK1 by down-regulating the GTPase RAC. However, CAPE alone has never
been clinically used, mainly due to its poor bioavailability (water-insolubility)
. Instead Bio 30, a CAPE-rich extract of NZ propolis available on the market
inexpensively, has been proven to be effective in blocking the growth of
several PAK1-dependent cancers such as pancreatic and breast cancers, NF
(neurofibromatosis) tumors, gliomas and melanomas in vivo (both xenograft
in mice and clinically), due to its high bioavailability and synergy between
CAPE and several other anti-cancer ingredients such as pinocembrin and galangin.
Thus, it would be quite reasonable to test the therapeutic effect of Bio
30 on human colon cancers in vivo, to begin with.

Besides Bio 30, Brazilian green propolis extract (GPE), which is based on ARC
(artepillin C), instead of CAPE, also inactivates PAK1 (Messerli, S. et al, 2009),
and suppresses the growth of colon cancers in vivo (Shimizu, K. et al, 2006),
although GPE is far more expensive than Bio 30. These findings altogether confirmed
that colon cancers indeed require PAK1 for their growth.

2009年11月11日水曜日

Gum Arabic: To be used for emulsifying EEP
(ethanol extracts of propolis) such as Bio 30

Gum Arabic (GA) is a natural "edible" water-soluble gum from Acacia that
has been used for a variety of purposes such as stamp glue and emulsifier.
Recently I got interested in GA mainly for the following two reasons: (1)
it can emulsify the water-insoluble ethanol extracts of propolis (EEP) such
as Bio 30, and (2) it appears to block the inflammation by inactivating
the transcription factor NF-kapperB (Rapnir, R. et al, 2008), down-stream
of the kinase PAK1, suggesting that it probably blocks the oncogenic PAK1
signaling. Besides GA is available inexpensively on the market, costing
less than US$50 per kg. To emulsify 6 ml of Bio 30 (costing around a dollar
daily) for an instance, you need only 6 g of GA powder (costing an additional
30 cents daily) in 6 ml of water (50% GA). So in my opinion, once NZ (or
Brazilian green/red) propolis is extracted with ethanol, this ethanol should
be replaced by 50% GA, instead of PG (propylene glycol). Furthermore, I
hope that GA and Bio 30 would work synergestically to suppress the growth
of a variety of PAK1-dependent cancers and NF (neurofibromatosis) tumors.

2009年11月3日火曜日

豪州でも久しぶりに「梅酒」作り!

「梅酒」作りの材料としては、青梅 1kg 当たり、氷砂糖200ー600 g、焼
酎1。8リットルが必要である。作り方の詳細については、下記のネット欄を参
照されたし。

http://kazuko-w.ld.infoseek.co.jp/ume_liquor.html

実は、ちょうど一ヶ月前に引っ越してきた新居の庭(両隣の家との境界近く)に、
以前の住人が残していったリンゴの若木など2、3の果樹の中に、どうやら梅の
実らしい物がなり始めた高さ3メートル近くの木を一本見つけた。引っ越し当初
は、木の葉の半分近くが植物ウイルスらしいものに感染したまま放置され、緑色
の木の葉の上に黄色や赤の(丸でヒトの腫瘍を連想させる)グロテスクなオデキ
状の物が至る所に見える惨々たる有様だった。その病気の木の葉を一か月かかっ
て、こまめに少しずつ取り除き、ようやく「手術」のかいあってか、(木全体の
サイズ=幅は半減したが)ほとんど健康そうな(細身の)木に回復した。

途端に、ずっと昔、東京の実家で、子供の頃(毎年、梅雨の時期になると)、亡
父と一緒に、兄妹3人で赤味がかった梅の実を収獲し、大きな瓶にどっさり砂糖
と梅の実を詰め込み、焼酎漬けしたのを、懐かしく思い出した。梅酒が美味しく
熟すには丸一年かかる。そこで、今年の12月(豪州メルボルンでは初夏に当た
る)には、久しぶり(半世紀ぶり)に自家製の梅酒を作ろうかと考えている。

この梅酒がちょうど熟する頃(来年の12月)には、目下台湾の台北大学で英文
学の教鞭にあたっている双子の娘のひとり(アイリス)が、一年3か月ぶりにメ
ルボルンに戻ってきて、私と一緒にこの新居で生活を始める予定になっている。
それを祝って、「梅酒で乾杯!」というのは、絶好のタイミングだと言えるだろ
う。。。

2009年10月21日水曜日

シシリー島の黄色い花 (Scotchbroom, Ginestra)

我が新居の近所をブラブラ散歩中、世にも不思議な花を見つけた。黄色い花だが、こ
の潅木には葉っぱが全く見当たらない。幹から枝が出て、そこから直接芽が出て、
非常に甘い芳香のする花が咲く(実は、このマグノリア=木蓮に似た香りが
私の鼻を真っ先に捉えた!)。

庭仕事に余念のない家のご主人(フィリップ老人)に尋ねたところ、母国シシリー島の
どこにでも生えている野生の潅木だそうだ。枝を2、3本もらったが、新居には花瓶など
備えてないので、(赤い)バケツに水をはって、そこに無造作に立てた(写真の前方。
後方=紫色の花は鉢植えのスペイン産のラベンダー)。繁殖には「種撒き」が必要で、
(枝をもらっても)插し木がきかないのは誠に残念だ。

この黄色い花の束を中庭に置くと、たちまちミツバチが数匹群がってきた。
すぐ隣にある (比較的芳香の弱い) ラベンダーなどには、目もくれなかった。
魅力の差が全く歴然としていた! 養蜂家としては、このエニシダ「Scotchbroom,
Ginestra」を我が家の園芸植物として最優先するほかないだろう。。。

イタリアのミラノ生まれの古い友人 (フランチェスカ) の話では、赤や紫の花を咲かせる
エニシダの品種もあるそうだ。黄色に加えて、赤や紫の品種も是非将来手に入れて、
我が庭に咲かせたいものである。

ところで雑談だが、西洋ではエニシダの枝から箒(ほうき)を作った。御伽話に出
てくる魔女(魔法使いのお婆さん)がまたがって空を飛ぶあの箒も、エニシダの
枝でできていると言われている。

2009年10月17日土曜日

丁公藤 (Erycibe obtusifolia Benth)

出典: http://www.moroo.com/uzokusou/nikki/114.html

感遇藥藤

南朝の鮮叔謙は、母が病気であった。鮮叔謙が、夜庭で神に祈っていると空中か
ら声が聞こえた。「この病気は丁公藤を入手して酒にするとよい薬になる」と声
は言った。鮮叔謙は、丁公藤を知っている人をさがしたが見当たらず、遂に宣都
に着いた。はるか遠くを見渡すと、一人の老人が木を切っているのを見つけた。
鮮叔謙は何に使うのかと訪ねると、「これは、病気の薬になる木で、丁公藤と言
うのだよ」と答えた。鮮叔謙は忽ち地面にひれふして土下座をして涙を流しなが
ら、母が病気である事を詳しく告げた。老人は、哀れんで鮮叔謙に丁公藤を与え、
酒にする方法を教えた。鮮叔謙は、伏し拝んでお礼を告げたが、老人の姿は無く
なっていた。教えてもらった方法で酒を作り、母に飲ませると病気は治った。

さて、この故事に登場する母親の難病については、詳しいことはいまだに不明で
あるが、私の科学的直感に頼るならば、恐らく癌かNFか痛風ではないかと思わ
れる。その科学的根拠を簡単に、下記に記そう。

最近、南京の中国薬科大学の薬理学研究室から大変面白い報告が発表された。丁
公藤の幹由来の「スコポリン」と呼ばれるクマリン配糖体が血管新生やリューマ
チなどの炎症反応を抑制するとともに、PAK1の下流にあるキナーゼ「ERK」
をも阻害することを発見した。ところで、血管新生や炎症反応にもPAK1が必
須であることを考え合わせると、スコポリンはどうやらPAK1を遮断している
可能性が極めて高い。従って、PAK1依存性の癌の7割以上、NFや痛風も将
来スコポリンで治療しうる可能性が出てきた。丁公藤の煎じ薬は、漢方で少なく
とも痛風の治療薬として古来使われてきた。

Int Immunopharmacol. 2009 Jul;9(7-8):859-69.

Scopolin isolated from Erycibe obtusifolia Benth stems suppresses adjuvant-induced rat arthritis by inhibiting inflammation and angiogenesis.

Pan R, Dai Y, Gao X, Xia Y.

Department of Pharmacology of Chinese Materia Medica, China Pharmaceutical University, 1 Shennong Road, Nanjing, Jiangsu Province, China.

Despite scopolin is a main coumarin constituent in the stems of Erycibe obtusifolia Benth, a herb drug that has long been utilized in traditional Chinese medicine for the treatment of rheumatoid arthritis, little information is available about the pharmacological activities of this compound. The present study was performed to investigate the anti-rheumatic effects of scopolin in adjuvant-induced arthritis (AIA) in rats, and explore the underlying mechanisms of action in views of anti-inflammatory and anti-angiogenic properties in the synovial tissues.

Scopolin (50, 100 mg/kg), injected intraperitoneally for 10 days from the onset of secondary response, significantly inhibited both inoculated and non-inoculated paw swelling as well as articular index scores in AIA. Meanwhile, the mean body weight of rats treated with scopolin was higher than that of model group. Rats treated with high dose of scopolin (100 mg/kg) preserved a nearly normal histological architecture of the joints and showed a significant reduction of the new blood vessels in the synovial tissues. Additionally, scopolin could reduce IL-6, VEGF and FGF-2 expressions in rat synovial tissues.

In conclusion, scopolin can reduce the clinical symptoms of rat AIA by inhibiting inflammation and angiogenesis, and this compound may be a potent agent for angiogenesis related diseases and can serve as a structural base for screening more potent synthetic analogs.

2009年10月16日金曜日

豪州独特の園芸植物1: 「BottleBrush」 (ブラシの木)

2週間ほど前に、メルボルンの郊外、西ブランズウックにある比較的広い庭を持
つ古いレンガづくりの平屋に引っ越した。園芸と養蜂を始めるためである。手初
めに、鉢植えの花を2、3、種苗園から手に入れて、中庭に植え付けた。その1
つが、豪州原産の真っ赤な花の咲く潅木で、「BottleBrush 」(ブラシの木) と
呼ばれている。花の形が珍しく、瓶洗いに使うブラシにそっくりだからだ。
春から秋にかけて、真っ赤なブラシ状の花を豪華に沢山咲かせる。実に壮観
である。木は大きく成長すれば、4メートル近くの高さにまでなる。色々な小鳥
が好んで飛んできて、このブラシの実をついばむ。南氷洋から吹き付けてく
る冷たい南風を防ぐために、南側の塀沿いに、この潅木(常緑樹)を殖やして
「防風林」代わりに植え込む積りである。

2009年10月6日火曜日

Elizabeth Blackburn: the First Aussie Woman who Won the Nobel Prize!

She was born in Hobart, Tasmania, in 1948, and graduated from Melbourne
Uni in 1970. Then she got her Ph. D. at University of Cambridge in 1975,
and started working at University of California at Berkeley. In 1982, she
and Jack Szotak at Harvard Medical School (Dana Farber Cancer Center)
jointly discovered the "telomeres" at the end of chromosomes in yeast.
In 1984, she and her Ph.D. student Carol Greider jointly discovered
a new enzyme called "Telomerase" which extends the telomeres.

This year Elizabeth Blackburn at UCSF, Carol Greider at Johns Hopkins University
and Jack Szotak at Harvard University (MGH) share the 2009 Nobel prize
for their pioneering work on Telomere and Telomerase.

Abnormal activation of Telomerase (or extension of telomeres) is a cause of
human(but not mouse) cancers which prevents normal cells from the senescence
(programmed cell death).

Interestingly human telomeres are far shorter than mouse counterparts. So
in general human normal cells are shorter-lived than mouse counterparts.
That is why it is very hard to establish "immortal" human normal cell lines.
For every time each cell divides, its telomers get shorter, and in the end
when its telomeres almost disappear, the cell is bound to die. Like a blasting
fuse linked to dynamite, shortening of telomeres leads to the burst of cells.

However, when human normal cells are transformed by tumor viruses such
as HPV, they are immortalized, because these viruses contain a gene for telomerase
which could extend the telomeres. The best known example is a human cervical
cancer cell line called "HeLa" which is transformed by HPV.

In principle, anti-telomerase drugs would be anti-cancer drugs potentially
useful for the therapy of human (but not mouse) cancers. In other words,
the anti-cancer efficacy of these anti-telomerase drugs could not be tested
in mouse cancer models. That might be a reason why the development of anti-telomerase drugs has been so slow.

Actually I have never met this Aussie-American scientist,
but I have spoken Carol Greider once as I invited her
among the speakers to our 1998 NY Academy of Sciences symposium
on Anti-cancer molecules which I organized at Rockefeller University.

2009年9月28日月曜日

海鞘の産物「STー2001」から強力なPAK阻害剤?

我々のCEPー1347をNF治療薬にするという計画は、「セファロン」の不協力で
実りそうもなかった。 そこで、もっと(千倍くらい)強力なPAK遮断剤を探し始めた。
CEPー1347は、前述したように、K252aに比較的長い側鎖をつけた誘導体だ。
ある知人から、K252aに似た化合物スタロスポーリン(ST)に同様な側鎖
をつけると、CEPー1347より20倍くらい強力なPAK遮断剤になること
を耳にした。 しかし、これはある会社内の開発途上物質で、我々には入手不可能
だった。 ここで得るべき大事なヒントは、STの誘導体で、PAK阻害のIC50が
1 nMくらいの物質を見つけ出して、こいつに同じような側鎖を施すことに
よって、「PAK特異的な」阻害剤に変え得るというアイディアであった。色々
スクリーニングしているうちに、それらしいものが海洋産物から見つかった。

もし、海鞘を常食にしている者は癌にならない、という伝説があったとしたら、そ
の由縁は、この誘導体のおかげかもしれない。なんとグアム島の沿岸に棲息する
特殊な海鞘から、STの3位に水酸基がついて化合物が、シドニーの海洋研究
所のピーター・シュップによって、2001年に発見された。特にPAKとPKC
というキナーゼを阻害する作用が強い。 しかしながら、PKC阻害剤は副作用が
強いので、常識的には制癌剤にはなりにくい。 そこでこの誘導体
(STー2001と命名)の9位に、(例えば、CEPー1347にあるような)
長い側鎖をつけることによって、特異的にPKC阻害活性を除き、
「PAK特異的な」阻害剤に変えるというアイディアが生まれた。

ここで更に欲を言えせてもらえば、9位に付ける側鎖として、アルキルアミン (例えば、
ヘキシルアミンなど) を選べば、このST誘導体の水溶性が増すと共に、細胞膜を通
過しやすくなるので、将来の臨床への応用に大いに寄与すると期待される。

さて、STー2001を化学的に合成することは可能であるが、収率が極めて低
いので、企業上実用的ではないと、ある有機化学の専門家にいわれた。従って、
この特殊な海鞘をグアム島から大量に採集して、それから微々たる量の
STー2001をせっせと精製する以外には、今のところ方法がないようだ。

もし、STの3位のみを特異的に水酸化する酵素(ST ヒドロキシラーゼ)を、この海鞘か
ら精製、あるいはその遺伝子をクローンすることが将来できれば、(グアム島ま
で出かけなくとも)実験室内で大量にSTー2001を酵素的に合成することが
できるだろう。そうすれば、夢のPAK遮断剤「STー3009」の実現も可能
になろう。

2002年に、富山県立大学生工研の古米保教授の研究室によって、
STの生合成に関与する14種類の遺伝子がクローンされた。面白いことには、
STの生合成経路を調べてみると、アミノ酸である「トリプトファン」2分子が
まず縮合してSTの芳香環を形成することがわかる。次に糖(ヘキソース)部分が
この芳香環に連結される。ところで、トリプトファンを水酸化する酵素 (TRP ヒドロキシラーゼ、
TPH)は、STの3位に相当するトリプトファン部分(5位)を水酸化することが知られている。
従って、理論的には、(既に結晶化もされている)TPHによって、STの3位を特異的に水酸化
し得る可能性もある。

そこで、STを大量に生合成する放線菌 (Streptomyces staurosporeus) 中に
TPH遺伝子(cDNA)を人工的に挿入して、その遺伝子の発現を IPTG(あるいは
熱ショック)などで効率良く誘導して、STー2001を大量に生合成できる新しい菌株
(STーTPH)を作り出すというアイディアが生まれつつある (実は、最近になって、
地球温暖化のためか、この特殊な海鞘がグアム島沿岸から姿を消しつつあるという
一種の危機に直面しているからだ!)。

ところで、TPHの働きには通常、2種類の異なる補酵素(3価の鉄イオンとBH4/
tetrahydrobiopterin) が必要である。 問題は、2番目の補酵素BH4が
(動物の細胞同様)バクテリアにも存在するかどうかである。 もし、存在しなければ、
バクテリアの培養液中に、この補酵素を補充しなければならない。理論的には、かくして
「STーTPH」菌株中でできたSTー2001を精製し、更に9位にヘキシルアミンを付加する
ために、アミノヘキサノールを加えて脱水縮合すれば、夢の水溶性PAK遮断剤「ST3009」が
誕生するはずである。

2009年9月24日木曜日

名言: 「貧乏人は麦を食え!」

「貧乏人は麦を食え!」という極めて率直な発言をしたのは、池田勇人(吉田内
閣の蔵相)だった。敗戦後まだ間もない1950年、米価が著しく高騰していた
頃の若き蔵相の英知だった(国民、その大部分が「貧民」、から激しい批判を浴
びたが)。勿論、貧乏人の先頭を行く我が家は率先して、米食から麦食(パン食)
に切り換えた。実は、我が家の家事を一手に預かる父(池田氏と同様京大出身で、
7年後輩)は、慢性胃潰瘍を長らく患っていたので、消化の悪いお米より、焼き
立ての食パンの方がずっとこなれ易いから、直ぐ先輩に同調したのだ。以来、我
が家は西洋式のパン食になった。だから、池田勇人の勇ましい発言に感謝せねば
ならない(10年後に、彼自ら首相になった!)。父の死後(ちょうど20年前
の今日!)、我が家(実家)は、また米食に戻ってしまったようだが(多分、安
い「古古米」が市場に溢れるような大変豊かなご時世になったからかもしれない)。

A Tiny Impact of Avastin on Cancer (Solid Tumors) and NF Therapy

Avastin, an anti-VEGF monoclonal antibody, was recently reported to improve
the hearing of NF2 (schwannoma) patients (4 in 10) in trials conducted by
a Mass General team. However, since Avastin from Genentech costs around
$15,000 monthly (around 500 times as much as Bio 30), only a few rich NF
patients can afford to keep taking it for the rest of their life.

Serious oncologists in general keep ignoring Avastin (among the most expensive
drugs), simply because it alone does not prolong the life span of cancer
patients. For just like Thalidomide, Avastin could reduce only the size
of solid tumors (by 50% or less) by blocking angiogenesis, but does not
block the metastasis which would kill the patients eventually. Actually
the old drug "Thalidomide" is far more inexpensive and safer than Avastin
for the same purpose (blocking angiogenesis).

NF Patients need an inexpensive "multi-functional" drug which blocks both
metastasis and growth (division) of cancer cells, in addition to angiogenesis,
as do propolis (Bio 30) and a few other anti-PAK drugs on the market such
as DPM (dipyridamole) and Ivermectin which pass BBB (blood brain barrier).

By the way, unlike Thalidomide, Avastin itself does not pass BBB, but just
sequesters VEGF from the blood circulation. Another limitation of Avastin
is the short-life of its therapeutic effect. For it is just a derivative
(humanized) antibody from mouse. In other words, it is similar to human
antibody, but not exactly the same. Therefore, soon or later, patients'
immune system would produce a neutralizing antibody which destroys Avastin.
Thus, the impact of Avastin on cancer/NF therapy would be rather tiny in
a long run, although its "price tag" is currently huge.

2009年9月18日金曜日

A Wisdom for the Potentiation of CAPE-based Propolis

NZ propolis is known to be the richest in CAPE (caffeic acid phenethyl ester)
, the major anti-cancer ingredient in propolis produced in Europe, Far East, and Oceania.
Although CAPE content in NZ propolis was reported to be 6-7 % (60-70 mg
per g of propolis), the highest around the world, for some unknown reason
the current NZ propolis on the market called "Bio 30" contains only one
fifth of the CAPE. Why? It remained a mystery until recently.

Is ethanol extraction of crude propolis insufficient for the full extraction
of CAPE, or Is CAPE converted to CA (caffeic acid) during the processing of crude propolis?

The crude propolis contains an enzyme called esterase which hydrolyzes CAPE
to CA. This enzyme would be activated as soon as the crude propolis is exposed
to water. The first step for the conventional processing of crude propolis
from bee matrix which collects propolis in bee hives is to suspend crude
propolis in water, which allows the wax (lipids) to float and the propolis
to be precipitated. The propolis is then air-dried and extracted by ethanol
which inactivate the esterase and solubilize the anti-cancer ingredients
such as CAPE and pinocembrin (PIN). Finally ethanol is replaced with PG
to make an alcohol-free liquid called Bio 30. Thus, it is most likely that
CAPE is converted to CA (by 70-80%!) in water during the first step.
We are getting a few lines of evidence supporting this notion, which would
solve this mystery around Bio 30.

In my opinion, this "wasteful" first step should be omitted to save the
precious CAPE and also wax which would solubilize CAPE and improve its bioavailability.
Instead, the crude propolis should be extracted directly from the matrix
of hives with ethanol to kill the esterase and extract both CAPE and wax.
By such a simplified procedure, the current Bio 30 (or any other CAPE-based
propolis such as Chinese red propolis ) could be further potentiated 5-10 times, without any extra cost.

Thus, this "tiny" change might revolutionalize the whole propolis industry
for the far more effective therapy of cancers and several other PAK-dependent
diseases.

2009年9月16日水曜日

全く期待外れの民主党「鳩山内閣」人事

一口に言えば、人材不足にもかかわらず、民間の専門家を一人も採用しようとし
ない、いわゆる旧態依然とした党人中心の「派閥人事」である。

小沢幹事長が裏で人事を工作している限り、民主的な適材適所の内閣は誕生しな
い。半年以内に内閣改造は必至である。改造しなければ、かつての細川内閣同様、
内閣の早期解散に追いやられるだろう。

有権者たちの「変化」への期待が大きかっただけに、失望感から来る怒り (逆風
) も殊更強まるだろう。このままだと、来春、桜が咲き始める時分に、鳩山内閣
(実質的には小沢氏の傀儡政権)が散り始める可能性がある。

花咲けど 実一つ結べず 風に散る

問題は、野党側に回わされた自民党が、寄付金 (企業献金) 不足のため、それま
でに空中分解 (破産) しているかどうかである。。。

私個人は、鳩山内閣による内政には余り期待できないが、「岡田外交」による独立国
「ニッポン」の欧米 (特に米国) との対等な外交関係の確立 (敗戦後半世紀以上続く
「米国追従外交」からの脱却) に期待したい。 岡田外相の健闘を心から祈る!

さて、公明党の今後の行方は? 賞味期限の切れた自民党との共闘にはもう旨味
が全くなくなった! それでは、新しい与党の民主党に接近して、社民党のごと
く「おこぼれ」を頂戴せんとするか、それとも共産党のごとく、どの政党とも共
闘せず、かたくなに、いわゆる「是々非々」(実際には「非々」ばかり)の立場を取るだろうか。。。
創価学会の信者たちは、公明党に一体何を望んでいるのだろうか?

2009年9月10日木曜日

DPM (ジピリダモール): 廉価で安全なPAK遮断剤

さて、PAK遮断剤「CEPー1347」やFK228は、RAS癌細胞やNF腫瘍 細胞に「接触阻害」(正常な細胞同士が接触すると、増殖を停止するというユニークな現象)を誘導し、腫瘍の増殖を効果的に抑制するが、まだ治験中で市販への道は、まだまだかなり遠い。そこで天然物ではなく「合成」薬品であるが、半世紀前から市販され、廉価かつ安全なPAK遮断剤を一つ、ここで紹介しよう。

NF2遺伝子産物「メルリン」がPAK阻害蛋白であることは前述した。そして、PAK阻害が「接触阻害」(いわゆる「ギャップ・ジャンクション」の形成) に必要であることも判明している。癌細胞に、この「接触阻害」能力が欠けているのは、PAKが異常に活性化しているからである。いいかえれば、抗癌蛋白「メルリン」によるPAK阻害が「接触阻害」に重要な役割を果たしていることが予想される。しかしながら、メルリン自身は細胞質蛋白で、貫通蛋白ではない。面白いことには、メルリンがある貫通蛋白に結合することが知られている。その蛋白は「CD44」と呼ばれ、ヒアルロン酸(HA)のレセプターであるが、RASを何らかのメカニズムで活性化することが、ドイツのイエナ大学の英国人ヘレン・モリソン のグループによって2007年に明らかにされた(1)。従って、メルリンがCD44の発癌性を抑えていることが予想される。実際、同じ年にペンシルバニア大学の中国出身の獣医、兪 勤 (ユー・キン) のグループは、メルリンがHAとCD44との結合を抑えていることを発見した(2)。

そこで、HAの生合成を抑える市販の化合物を物色しているうちに、最近「DPM」という薬剤に突き当たった。この阻害剤は、HAによるCD44の活性化を遮断するので、抗癌蛋白メルリンと同様、RASによる癌化およびメルリンの不全により発症するNF2腫瘍 (シュワノーマやメニンジオーマ) の増殖を抑えるわけで、これらの癌やNFの特効薬として、有効である可能性がにわかに浮上してきた。

DPMは1950年ごろ、ドイツの製薬会社「ベーリンガー」によって開発された薬で、血小板の凝固を抑える機能があるため、脳溢血などの治療薬として半世紀以上、世界中で広く使用されてきた安全かつ安価な医薬品である。ところが1985年になって、英国のエリザベス・ローデスのグループによって「DPMがメラノーマにも効く」という治験結果が報告された(3)。つまり、DPMには抗癌作用もあるというわけである。さらに1989年には、もう1つの新たな機能、薬剤耐性に関与するMDRというポンプATPase (薬剤を細胞外へ汲み出す酵素) を阻害する作用が、DPMにあることがイスラエルのアブナー・ラムー夫妻によって発見された。従って、薬剤耐性になった癌患者の治療にも役立つ可能性が出てきた。DPMには、PAK遮断剤と同様、癌の転移や血管新生を抑える作用もあることがわかっている。

さて、ごく最近になって、このDPMの薬理作用に大変面白い展開が起こった。私の旧友で、ドイツのミュンスター大学で教授をやっているピーター・プレム はHA合成酵素 (HAS)に関する研究の専門家だが、彼にHAS阻害剤について、問い合わせたところ、なんと「DPMが市販されている薬剤のうちで、最も強いHAS阻害剤だ」という返事がすぐ戻ってきた。実は2004年に、彼の友人であるUKEのウド・シューマッハーとの共同研究で、DPMがHASを阻害することを、偶然見つけたのだそうだ(4)。こうして、DPMが発癌性のRASーPAKシグナル経路を抑えることが判明したわけだ。早速我々は、DPMがNF1癌であるMPNST細胞やNF2腫瘍であるシュワノーマ細胞の増殖を抑えることを実際に確認した。


参考文献

1. Orian-Rousseau, V., Morrison, H., Matzke, A., Kastilan, T. et al. Hepatocyte growth factor-induced Ras activation requires ERM proteins linked to both CD44v6 and F-actin. Mol Biol Cell. 2007, 18, 76-83

2. Bai, Y., Liu, YJ., Wang, H., Xu, Y. et al. Inhibition of the hyaluronan-CD44
interaction by merlin contributes to the tumor-suppressor activity of merlin. Oncogene. 2007, 26, 836-50.

3. Rhodes, E., Misch, K., Edwards, J., Jarrett, P. Dipyridamole for treatment of
melanoma. Lancet. 1985, 1(8430), 693.

4. Prehm, P., Schumacher, U. Inhibition of hyaluronan export from human fibroblasts by inhibitors of multidrug resistance transporters. Biochem Pharmacol. 2004, 68, 1401-10.

2009年9月8日火曜日

岡田外相へ進言: 「大使」たるべき人物の人選

一般論

大使の任命について、一言したい。私は豪州に21年以上永住しているが、駐豪
日本大使の名前を一人も知らない。なぜかといえば、しょっちゅう(数年毎に目
まぐるしく)変わるからだ。さらに、我々(永住日本人)の利益とは、ほとんど
無関係の仕事をしているからだ。我々と日本大使館(厳密にいえば、メルボルン
の領事館)と関係は、10年に一度、長期パスポートの延長と、最近は(総選挙
と参議院選挙)の在外投票にタッチすることだけだ。実際には、これらの仕事を
しているのは、大使や領事自身ではなく、その下で長らく現地で働く事務官たち
である。

そこで、提案したいのは、大使や領事も、その任地で長らく働き、現地の人々や
文化に精通している日本人から選ぶべきであるということだ。

一例をあげれば、インド駐在のニュージーランド大使(高等弁務官)となったエ
ドモント・ヒラリー卿(1919ー2008)だ。 彼は1953年に、シェル
パのリーダー(テンジン)と共にエベレスト初登頂に成功して以来、現地ネパー
ルの貧しいシェルパ民族のために、病院や学校を建設する慈善事業に長らく携わっ
た。こういう人物が大使に任命されるべきで、現地の実情を全く知らない高級
(外務)官僚が(単なる「腰かけ」で)大使として、数年だけ現地に滞在してい
ても、何の実益にもならない(国税の無駄使いに過ぎない!)。  


国連代表の人選

従来の自民党による「米国追従」外交を打破し、「米国と対等」で我が国の外交
を推進するためには、「岡田外交」に相応しい新しい国連大使(代表)を緊急に
抜擢する必要がある。私は、高須(現)国連代表にかわって、リベラルな天木直
人氏(元レバノン駐在大使)を国連代表に起用することを、強く推薦したい。

天木さんは、2003年に小泉政権が米国のブッシュ政権に追従して、イラク侵
略戦争に加担しようとした瞬間、先見の明をもって、(現職の外交官で)ただ独
り、勇気をもって、小泉政権のイラク戦争への加担に対して、親書ではっきり
「ノー」を表明して、大使辞任を強要された、筋金入りのいわば「独立外交官」
である。彼は1969年に京大から、外務省に入省した(東大出身の高須さんと
偶然にも同期)。彼の外交官としての気概は、著書「さらば、外務省」に如実に
述べられている。

彼の豊かな経験と気概によって、日本の「平和外交」を国連という国際舞台で、
強く推進してもらおうではないか! リベラルな現国連事務総長(潘 基文)の
片腕として、地球から戦争をなくし、さらに地球の温暖化を防ぎ、世界全体の環
境を改善するための政策(呼びかけ)を実行してもらいましょう! 

国際外交をいつまでも欧米人たちだけに任せておかず、今後は我々アジア人がイ
ニシアチブをとって世界の外交を動かしていかなければ、世界の真の平和は望め
ない!                   丸田  浩(66、メルボルン永住)

Therapeutic Hope for TSC (Tuberous Sclerosis) Patients

A complex disease called TSC (Tuberous Sclerosis) is caused by deletion or dysfunction of either TSC1 or TSC2 genes. Both gene products are tumor suppressors which form a complex that normally serves as a GAP (GTPase activating protein=attenuator) of a GTPase /G protein called Rheb. Thus, in TSC patients, Rheb is abnormally activated (becoming oncogenic!), and then activates another kinase "TOR" that in turn activates the potentially oncogenic S6 kinase. Thus, in many cases TSC is associated with a variety of tumors, which mainly develop in brain, spleen and kidney. Since these tumors very often develop in a quite early stage of life, most of TSC patients' life span is significantly shorter than non-TSC people's. Unfortunately, so far no effective (FDA-approved) therapeutic is available on the market. However, since "TOR" (target of rapamycin) could be effectively inhibited by the antibiotic "Rapamycin" or its more potent derivative "CCI-779" in rodent TSC models (1), in the future (in several years, hopefully) when clinical trials of "CCI-779" are successfully completed for the approval by FDA, the anti-cancer drug "CCI-779" will become available for the treatment of TSC and other formidable tumors.

Of course, many TSC patients, in particular parents of TSC children, often asked me if there is any effective "alternative" (non-FDA-approved) medicines or natural products which would be helpful for improving the conditions of TSC patients. In the past, until very recently, sadly I had to answer nothing but "No". However, eventually this year, I would be able to say to these TSC patients, "YES, YOU CAN have a specific alternative medication. There are a few potentially effective natural anti-PAK products on the market which would be helpful for TSC and other formidable diseases such as NF (neurofibromatosis) ". I will tell you how the situation has been dramatically changed. It has nothing to do with Barack Obama's new administration at the White House, but a few dedicated bio-medical scientists discovered something useful for the therapy of TSC patients.

The kinase "PAK" is essential for the growth of more than 70% of cancers and NF tumors, and therefore we have developed a series of anti-PAK drugs (synthetic chemical compounds) and also identified several anti-PAK natural products available on the market inexpensively, such as NZ propolis extract called "Bio 30", curcumin, ivermectin, and vitamin K2 (menaquinone-7) from "Natto", for these PAK-dependent tumors.

Interestingly, this year, a few scientific groups began providing an evidence suggesting that TSC tumors appear to be among these PAK-dependent tumors. First of all, curcumin, a yellow spice in Indian curry, inhibits PAK directly (2). Secondly, curcumin blocks the Raptor-TOR interaction (3). Raptor is a protein essential for the oncogenicity of TOR, and therefore, curcumin could block the oncogenic TOR signaling in TSC tumors. However, the bioavailability of curcumin alone is very poor (mainly due to water-insolubility), and without liposome or CD (cyclodextrin) which would solubilize curcumin, curcumin alone won't be effective for clinical application.

However, Bio 30, which is a water-miscible extract, and "Natto", a fermented sticky soybean product, which has been a traditional Japanese cuisine for a thousand years, would be effective for the treatment of both NF and TSC tumors as well as several PAK-dependent cancers such as pancreatic, colon, breast and prostate cancers as well as glioma, melanoma and MM (multiple-myeloma).

Also around two years ago, it was found that PAK inactivates the tumor suppressor "FOXO" that normally suppresses "Raptor" gene (4-6). In other words, these natural anti-PAK products on the market inactivate "TOR" by down-regulating "Raptor" in TSC tumors. Thus, I am now pretty convinced that TSC tumors could be treated with these anti-PAK products, as NF tumors.

Currently more than 150 NF patients are in my trials of Bio 30 world-wide, and we would welcome any TSC patients to join my trials. The recommended daily dose of Bio 30 (alcohol-free liquid) is 1 ml per 10 kg of body weight, and it costs only a dollar or so for the daily treatment. It causes no side effect, except for an allergic skin reaction which occurs only in 1% of population, due to its CAPE (caffeic acid phenethyl ester) which is the major anti-PAK ingredient (7).

If you wish to order Bio 30 (25 ml bottles) at NF/TSC discount price by joining my trials, please contact directly Annette Rea of Manuka Health, NZ: Annette@manukahealth.co.nz


References:

1. Lee, L., Sudentas, P., Donohue, B., Asrican, K. et al. Efficacy of a rapamycin analog (CCI-779) and IFN-gamma in tuberous sclerosis mouse models. Genes Chromosomes Cancer. 2005, 42, 213-27.

2. Cai, XZ., Wang, J., Li, XD., Wang, GL.et al. Curcumin suppresses proliferation and invasion in human gastric cancer cells by downregulation of PAK1 activity and cyclin D1 expression. Cancer Biol Ther. 2009, 8, 1360-8.

3. Beevers, C., Chen, L., Liu, L., Luo, Y. et al. Curcumin disrupts the Mammalian target of rapamycin-raptor complex. Cancer Res. 2009, 69, 1000-8.

4. Maruta, H. An innovated approach to in vivo screening for the major anti-cancer drugs. In “Horizons in Cancer Research” 2010: 41 (ed. Morrison, EP), pp249-59, Nova Science Publishers.

5. Jia, K., Chen, D., Riddle, D. The TOR pathway interacts with the insulin signaling pathway to regulate C. elegans larval development, metabolism and life span. Development. 2004, 131, 3897-906.

6. Southgate, R., Neill, B., Prelovsek, O., El-Osta, A. et al. FOXO regulates the expression of 4E-BP1 and inhibits mTOR signaling in mammalian skeletal muscle. J Biol Chem. 2007. 282, 21176-86.

7. Demestre, M. Messerli, S., Celli, N., Shahhossini, M. et al. CAPE (Caffeic Acid Phenethyl Ester)-based Propolis Extract (Bio 30) Suppresses the Growth of Human Neurofibromatosis(NF) Tumor Xenografts in Mice. Phytother. Res. 2009, 23, 226-230.


The corresponding author:

Prof. Hiroshi Maruta, Manager
NPO “NF CURE Japan”,
Melbourne, Australia.

The former head of Tumor Suppressor Lab.
Ludwig Institute for Cancer Research
(Melbourne Branch)

E-mail: maruta19420@mac.com or julie8860@gmail.com

2009年9月2日水曜日

Propolis is Good for Diabetes, Why?

Propolis in general, whether it is based on CAPE or ARC (Artepillin C),
has been known to be useful for the treatment of diabetes (both types 1
and 2). However, it remained unclear for a while how propolis could reduce
the blood sugar levels. Well, in 2007, a Korean group led by Hyeon Soo Kim
at Korean University Medical School in Seoul found that CAPE activates the
kinase AMPK somehow, leading to the reduction of blood sugar level by activating
a glucose transporter called "GLUT-4". Then, I wonder why ARC-based propolis
such as Brazilian green propolis is also effective in reducing the blood
sugar levels. I postulate that ARC must be responsible for this anti-diabetic
action, because nothing else in this propolis is good for the treatment
of cancers, NF, TSC and several other PAK-dependent diseases.

What is the most common biological property shared between CAPE and ARC?
Both block selectively the oncogenic kinase PAK.

Interestingly, there are several other natural products that block the kinase
PAK as well as activate the kinase AMPK, as does CAPE: curcumin, berberine,
resveratrol (trans R3), salidroside, and triterpenoids from bitter melon,
Moreover, both PAK inactivation and AMPK activation lead to the same outcome,
activation of the tumor suppressor "FOXO" which is essential for both the
activation of Hsp16 gene and the longevity. So I would hypothesize that
these natural products activate AMPK most likely by inactivating PAK which
I think normally inactivates AMPK somehow, proposing a new unified formula:

anti-PAK drugs=AMPK activators=FOXO activators=Hsp16 activators= "elixirs"


In other words, the best (quickest and least expensive) in vivo drug screening
system to explore the anti-PAK drugs (=elixirs) would be to monitor the
GFP expression in C. elegans (strain CL2070) which carries the Hsp16-GFP
fusion gene, that is rapidly activated shortly after heat-shock treatment,
only if PAK is blocked.

2009年8月23日日曜日

民主主義の夜明けが怒涛のごとく押し寄せている!

そういう形容がピッタリの総選挙直前の我々「進歩派」の心況である。
半世紀も続いた自民党による独裁「万年」政治にとうとう終止符が打たれる時期
が到来しようとしている。そう思っているのは、私独りではなかろう。しかしな
がら、15年ほど前に、ハレーすい星のごとく現れ、瞬く間に消えていった細川政権の
ことを思い出すと、油断は禁物だ! 来たるべき民主党(鳩山)政権が、約束し
た改革をきちんと実行しないと、あの細川政権の二の舞を演じる可能性はまだ十
分ある。

自民党という「怪物」を再生できぬよう土台から徹底的に崩すには、マニフェス
トの実行により、天下り官僚体制を根刮ぎしなければならない。政治や経済、医
療や教育を我々民衆の手に取り戻せねばならない。官僚主義から民主主義への変
換をできるだけ断行しなければ、鳩山政権は短命に終り、自民党政権が(中身自
体はそのままで)外見だけを変えて、また復活するだろう。村山(連立)政権が
その前例だ。それを絶対に許してならない!

2009年8月21日金曜日

Perseverance: True Voices of Cancer Survivors
by Carolyn Rubenstein(2009)

In general cancers are among diseases for old people. However, many young
people including children also suffer from some cancers such as leukemia
and glioma as well as NF and TSC tumors.

Like Lans Armstrong, the famous cyclist who overcame his formidable cancer
and won Tour de France seven times in row, so many other youngsters overcome
their cancers in their own way, and keep pursing their career. Carolyn Rubenstein
(24), a Ph.D. student at Harvard, published recently her inspiring story
about these 20 young brave and lucky cancer survivors who overcome their
fear caused by their life-threatening cancers in their childhood.

Anti-cancer drugs work only when cancer patients fight against this fear,
with a "positive" thinking. After all, there are a few endogenous anti-cancer
signal substances in our brain which are activated only when our mind looks
forward to the survival. Carolyn, who is currently studying clinical psychology,
has learned such an endogenous healing power, since she established a non-profit
organization "CCC" to support children with cancers, a decade ago.

Reading through this book, I hope every cancer patient, young or old, will
get both courage and energy for surviving his or her own cancer.

This book reminds me of at least two old friends of mine. One is my old German
Ph.D student who also overcame his leukemia a few decades ago, and is now
working in Sweden as a senior biomedical scientist. The other is Tom, a
professor at Yale as well as a Boston Marathorner and mountain climber,
who used to work at NIH for a few years in early 1970s, and found a "cofactor"
of a mysterious single-headed myosin from a soil amoeba. A year after he
returned to Harvard Med School, I started unveiling the very identity of
this cofactor which turned out to be a kinase which phosphorylates the heavy
chain of this myosin. Later this kinase was called "PAK". PAK is essential
for the growth of more than 70% of all human cancers as well as NF and TSC
tumors. Without his discovery of this amoeba cofactor, the advance in our
R and D of anti-cancer=anti-PAK drugs would have been significantly delayed.

Back to mid-1980s, Tom was suddenly found to suffer from a deadly cancer.
However, he was firmly determined to fight back with this cancer, and by
early summer of 1988, he was fully recovered from the cancer, and challenged
successfully the Himalayas with his wife and their children together, which
had been their life-long dream. For this special reason, I am planning to
publish my Japanese translation of this inspiring book with his family photo
in the Himalayas as a cover, hopefully to share these great "fighting spirits"
with youngsters as well as oldsters in my home country.

The end of this month, a big change will be expected in Japanese politics.
The bloody conservative party, which has cancerously ruled the Japanese
government for five decades, will be defeated at the coming general election
(August 30), and Democrats will grab the very first chance of ruling our
government. Our "perseverance" is about to be paid off at last!

2009年8月18日火曜日

短篇小説「日照権」

終身刑の囚人が、市の郊外にある我々の刑務所へ護送されてきた。小柄な細見の
老人だった。多少前こごみ気味だったが、しっかりした足取りで護送車から出て
きた。髪はほとんど真っ白だった。罪状は、自分が引っ越しを予定していた新居
の広い裏庭のすぐ真ん前(北側)にある2階建てのアパートを爆破し、一人の死
傷者を出したとのこと。我々が住むこの大陸は南半球にあるので、太陽は東から
出て、北側を回り、西に沈む。

6週間ほど前、老人はふいに一通の手紙を市役所から受け取ったそうだ。
「マッコル街8番地にあるアパートを3階建てに改造する工事の申請書を2週間
前に受け取った。もし、この工事に反対する者があれば、明日までに文書をもっ
て、市役所に異議を申し出ること」という内容だった。
老人には、マッコル街の拡張工事と自分が一体どんな関係にあるのかがすぐ呑み込め
なかったが、しばらくして、6週間先に引っ越し予定の自分の新居の直ぐ裏
にあるアパートが、「マッコル街8番地」に当たることに気づいた。老人はかん
かんに怒った。あの陽当りのよい広い庭で、ミツバチの好きな芳香の草花を植え、
養蜂を始めることを余生の最大の楽しみにしていたからだ。3階建てのアパート
は、確実に自分の新居の庭を暗くし、さらに新居の屋根に取り付ける予定の(湯
沸かしに利用する)ソーラーパネルの効率を損うのは明らかだった。早速、老人
は市役所宛てに、この拡張工事計画を却下するよう、要請書を提出した。

間もなく、老人の下宿(仮住い)に市役所からの返事が届いた。
「この工事に反対するのは、あなた独りなので、市はこの工事計画を承認するこ
とに決定した」
そこで、老人は、新居への引っ越しを取り止め、この新居の売却をキャンセルし
て、現在の住人(家主)へ「担保」として前払した350万円分を取り戻すため、市
役所あるいはアパートを経営する不動産会社に対して、同額の慰謝料を支払うよう、強く
要求した。しかし、その要求もあっさり却下された。そこで、老人はある計画を
立て始めた。マッコル街には、現在アパート群が6番地、8番地、10番地、12番地と
続けて並んでいた。いずれも2階建てだった。8番地が3階建てになれば、残り
のアパートも遅かれ早かれ3階建てに拡張されるのは、明らかだった。日照権の
侵害はさらに深刻にある。
「拡張工事に反対して、もし、8番地のアパートを爆破したら、どうなるだろう
か?」 
老人は「短気は損気である」ことを良く承知していた。しかし、手遅れになる前
に、やるべきことはしなくてはならない!
老人はある日、そのアパートにダイマイトを仕掛けた。アパートの住民たちに退
去を通告してから間もなく、アパートが木っ端微塵に吹き飛んだ。ところが、
不幸にして、難聴の老婆が一人逃げ遅れて、壊れたアパートの下敷きに
なって死んだ。老人は、家を買うために自分が一生をかけて貯めたお金を全部、
その遺族に支払った。老人には、もはや家を買う必要がなくなったからだ。

老人は我々の刑務所で、その一生を平和に過ごす機会を得た。この国には、死刑
が廃止されてから、もう何十年にもなる。温和しい老人は、やがて、刑務所内の
中庭にある花壇の管理を託された。数年後には、念願の養蜂を始めることも許さ
れた。老人は、この新しい「安住の地」で、長い「さまよえるオランダ人」の旅
をやっと閉じることができた。やがて、この老人の指導下に我々の刑務所の囚人
たちが生産する良質かつ廉価な蜂蜜やプロポリスが世界中に普及する日が訪れる
かもしれない。

私もこの刑務所の住人である。この刑務所はその昔、死刑囚を主に監禁していた施
設である。歴史的に有名な「ネッド・ケリー 」という英雄もここで最期を遂げた
といわれている。1967年に死刑制度が全廃されて以来、ここは終身刑囚人た
ちの安住の地になった。私もここの生活はもう長い。既に10年近くになる。囚
人の中には、ごく稀れだが、無実が晴れて、20数年ぶりに「しゃば」へ解放され
ていく者もいる。その時は刑務所あげてのお祭り騒ぎだ。囚人ばかりではなく、
監視たちも一緒になって、なごりを惜しんで送別会をやる。しかし、それは10
年に一度あるかないかの稀れな出来事である。大部分はここの独房でさびしく一
生を終える運命にある。私の独房番号は13番、なんと隣の独房14番に、あの
新参の老人が入居することになった。その奥の独房15番はまだ空き家のままだっ
た。そこで老人への歓迎会を私が引き受けることになった。老人はユーモアの持
ち主で、我々が着ている横縞の囚人服は、丸で「ミツバチ」の服装のようだと形
容した。そこで、有名なミツバチ「マーヤ」にちなんで、老人を以後「マーヤ」
という愛称で呼ぶことにした。

2009年8月16日日曜日

Let’s Protect Our “Solar Energy Rights” from the “Greedy” Real Estate Company!

If Moreland City Council in Melbourne fails to reject this “outrageous” construction plan on 8 McColl Court, Brunswick West, I shall cancel my purchase of the property just behind this planned “three-storey” apartment, and ask the council or the company Metro Corp (McColl) PTY to reimburse me $42,800 that I have already paid to the vendor as a 10% deposit. In this case, the vendor will get none of the remaining 90% from me on October 2. The reason is crystal-clear. The northern (sunny) garden of this property overshadowed by this “ridiculous” three-storey apartment, that would severely undermine our “solar energy rights”, would no longer suit to my professional gardening and bee-keeping career for the rest of my life.

In short, only if the council wisely rejects this construction plan in time, namely by the end of this August, I shall move into this property as scheduled.

2009年8月13日木曜日

A New Functional Domain of NF1 Molecule
at the N-terminal Half (1-1163)

This summer Yoel Kloog's group at Tel Aviv University in Israel found that NF1 gene
product of 2818 amino acids contains an additional functional domain (the first 1163
amino acids) that blocks the oncogenic RAC-PAK-LIMK signaling pathway.

Since the C-terminal RAS GAP domain of NF1 gene product attenuates
normal RAS, and blocks the same potentially oncogenic signaling pathway,
a loss-of-function mutation in either N-terminal (apparently RAC GAP) or
C-terminal (RAS GAP) half causes the abnormal activation of this PAK-LIMK
cascade, eventually transforming normal cells. It still remains to be clarified
why this NF1 molecule contains two separate but functionally similar (redundant)
domains.

It is of great interest to note that CAPE (caffeic acid phenethyl ester)
, the major anti-cancer ingredient in the NZ Propolis extract "Bio 30",
inexpensively available on the market, appears to do the exactly same job
as this N-terminal half of NF1 molecule.



Mol Cell Neurosci. 2009 Aug 7.

The Pre-GAP-related Domain of Neurofibromin Regulates Cell Migration through
the LIM Kinase/Cofilin Pathway.

Starinsky-Elbaz S, Faigenbloom L, Friedman E, Stein R, Kloog, Yoel*
* kloog@post.tau.ac.il

Department of Neurobiology, George S. Wise Faculty of Life Sciences, Tel
Aviv University, 69978 Tel Aviv, Israel.

2009年8月11日火曜日

TSC (結節性硬化症 ) 腫瘍に対する治療薬の開発(最新情報)

しばしば腫瘍を伴う結節性硬化症 (TSC) は、抗癌遺伝子 TSC1あるいは TSC2の欠損あるいは不全のために発生する稀少難病である。脳内あるいは腎臓に腫瘍が発生するケースが多い。稀少難病中、NFについで患者数が多く、世界的には百万人近いTSC患者が、NFと同様、有効な治療薬が未だ市販されぬまま、幼時から症状に苦しみ、多くの場合、短命で死亡する。

しかしながら、TSCに有効であると思われる治療薬が2、3目下開発途上(臨床テスト中)にはある。要は「時間の問題」である。市販ができるだけ早期に実現されれば、それだけ多くの幼いTSC患者の命を救うことができるわけだ。さて、TSC1蛋白とTSC2蛋白は、通常お互いに結合し合って、複合体を形成している。その機能は主に、G蛋白の一種である「Rheb」に対する「GAP」役を演じている ( 1 )。つまり、発癌性の「Rheb」の細胞増殖促進機能を、通常は抑制している。従って、TSC複合体の機能が損なわれているTSC腫瘍では、「Rheb」が異常に活性化されている。それが腫瘍化の主因である。それでは、「Rheb」は分子レベルで一体何をしているのだろうか?

数年前の欧米での研究結果から、G蛋白「Rheb」は、「TOR」という蛋白キナーゼを活性化する機能を持つことがわかっている ( 2 )。「TOR」とは、「Target of Rapamycin」の略で、「ラパマイシン」と呼ばれる制癌性の抗生物質によって阻害される キナーゼである。このキナーゼもPAKと同様、チロシンキナーゼではなく、蛋白基質のセリンあるいはスレオニン残基を燐酸化する酵素である。いいかえれば、TSC腫瘍の増殖は、「ラパマイシン」やその誘導体「CCIー779」などによって、少なくとも動物実験レベルで、ほとんど完全に抑制されることが数年前に確認されている ( 3 )。問題はそれが市販されるまでには、多分あと数年ほどかかりそうであるということである。現行の臨床実験は一般に、余りにも時間とお金がかかり過ぎている!

そこで、「ラパマイシン」類が市販されるまで、TSC治療に使用しうる代替薬は、この世に一体ないのだろうか? TSCに苦しむ子供を抱え込む多くの両親たちからの、そのような切実な訴えに応えて、最近、その周辺を文献上で調べてみた。すると、「無きにしもあらず」である! 要するに、「TOR」の機能を何とか抑えることができさえすれば、TSCは治療しうるわけである。そこで、最近注目を浴びているのが「Raptor」という蛋白である。実は、この蛋白が「TOR」の活性化に必須であることが、2002年に神戸大学の米澤一仁教授の研究室によって、明らかにされた ( 4 )。それだけではない! この「Raptor」の発現や「RaptorーTOR」相互作用には、なんと「PAK」が必須であるらしいことが、ごく最近 (今年になって) わかった。中国東北地方の中心都市の瀋陽にある中国医科大学や米国のルイジアナ州立大学の研究グループによって、インドカレーでおなじみのウコンの抗癌主成分「クルクミン」がPAKを遮断する( 5)とともに、「Raptor」を失活させる(6)ということが、独立に発見されたのだ! いいかえれば、クルクミンやNZ(ニュージーランド)産プロポリス「Bio 30」などの市販された天然物でPAKを遮断すれば、「RAS癌」やNF腫瘍と同様、TSC腫瘍の増殖も抑えることができるというわけだ!

こうして、2005年前後から開始した「廉価に市販されている天然物の中から、PAKを遮断する食物や生薬などを物色してみよう」という我々のユニークな研究努力が、つい最近になって、いよいよ「実用化」に向かって、実り始めてきたわけである! なお、この画期的な発見への糸口を作った米澤 一仁教授(神戸大学・バイオシグナル研究センター) は惜しむらくも、49才 の若さで急死されたそうである (2005年夏)。将来が大いに期待されていただけに、誠に残念である。

ただし、クルクミンは水に不溶性なので、胃腸からの吸収がひどく悪く、臨床的には、そのままでは実用性に乏しいが、リポソームやCD(環状オリゴ糖、シクロデクストリン) などで効率良くクルクミンを可溶化できれば、きっと近い将来、治療にも有効になるだろう。

参考文献

1. Manning BD, Cantley LC. Rheb fills a GAP between TSC and TOR. Trends Biochem
Sci. 2003, 28, 573-6.

2. Saucedo LJ, Gao X, Chiarelli DA, Li L, Pan D, Edgar BA. Rheb promotes cell
growth as a component of the insulin/TOR signalling network. Nat Cell Biol.
2003, 5, 566-71.

3. Kenerson H, Dundon TA, Yeung RS. Effects of rapamycin in the Eker rat model of
tuberous sclerosis complex. Pediatr Res. 2005, 57, 67-75.

4. Hara K, Maruki Y, Long X, Yoshino K, et al. Raptor, a binding partner of target
of rapamycin (TOR), mediates TOR action. Cell. 2002, 110, 177-89.

5. Cai XZ, Wang J, Li XD, Wang GL, et al. Curcumin suppresses proliferation and
invasion in human gastric cancer cells by downregulation of PAK1 activity and
cyclin D1 expression. Cancer Biol Ther. 2009, 8, in press.

6. Beevers CS, Chen L, Liu L, Luo Y, et al. Curcumin disrupts the Mammalian target
of rapamycin-raptor complex. Cancer Res. 2009, 69, 1000-8.


文責; 丸田 浩 (薬学博士)
NPO「NF CURE Japan」マネジャー
元ルードビッヒ癌研 (メルボルン支部) 制癌剤開発部長
連絡先: maruta19420@mac.com


Genes Chromosomes Cancer. 2005 , 42, 213-27.

Efficacy of a rapamycin analog (CCI-779) and IFN-gamma in tuberous sclerosis mouse models.

Lee L, Sudentas P, Donohue B, Asrican K, Worku A, Walker V, Sun Y, Schmidt K, Albert MS, El-Hashemite N, Lader AS, Onda H, Zhang H, Kwiatkowski DJ, Dabora SL.

Division of Hematology, Brigham and Women's Hospital, Boston, MA 02115, USA.


Tuberous sclerosis complex (TSC) is a familial tumor disorder for which there is no effective medical therapy. Disease-causing mutations in the TSC1 or TSC2 gene lead to increased mammalian target of rapamycin (mTOR) kinase activity in the conserved mTOR signaling pathway, which regulates nutrient uptake, cell growth, and protein translation. The normal function of TSC1 and TSC2 gene products is to form a complex that reduces mTOR kinase activity. Thus, mTOR kinase inhibition may be a useful targeted therapeutic approach. Elevated interferon-gamma (IFN-gamma) expression is associated with decreased severity of kidney tumors in TSC patients and mouse models; therefore, IFN-gamma also has therapeutic potential.

We studied cohorts of Tsc2+/- mice and a novel mouse model of Tsc2-null tumors in order to evaluate the efficacy of targeted therapy for TSC. We found that treatment with either an mTOR kinase inhibitor (CCI-779, a rapamycin analog) or with IFN-gamma reduced the severity of TSC-related disease without significant toxicity. These results constitute definitive preclinical data that justify proceeding with clinical trials using these agents in selected patients with TSC and related disorders.

2009年8月4日火曜日

Polyphyllin D: A Potent Anti-Cancer Saponin
of the Chinese Herb "Paris polyphylla"

In 2001, a Chinese group led by Kwok-Pui Fung at Hong Kong University developed
an improved method for the chemical synthesis of a saponin called "Polyphyllin
D" (PPD), the major anti-cancer ingredient in a Chinese herb (root of Paris
polyphylla).

In 2005, his group reported that PPD solubilized in gamma CD (cyclodextrin)
suppresses almost completely the growth of breast cancer xonografts in mice
at the daily dose of 2-3 mg/kg. Interestingly, most of cancers sensitive to
this saponin appear to be among PAK1-dependent cancers such as breast,
prostate, liver, colon, stomach, pancreatic and lung cancers.

This year, two Chinese groups in China and US jointly revealed a molecular
mechanism underlying the anti-cancer action of this saponin (PPD). Just like
FK228, PPD inhibits the expression of VEGF and HIF-1 alpha. VEGF is essential
for tumor-induced angiogenesis. HIF-1 alpha (hif-1) is a hypoxia (oxygen
depletion)-inducible factor which is required for the expression of VEGF.
Furthermore, expression of hif-1 requires the oncogenic kinase PAK1 which
inactivates "FOXO" that is essential for the longevity. In other words, PPD
would contribute to the longevity, most likely by blocking PAK1.

The reason is as follows. Recently it was found that down-regulation of hif-1
prolongs the life span of a tiny nematode called C. elegans through "FOXO",
indicating that PPD, which down-regulates hif-1, would prolong the life-span at
least in two distinct ways through "FOXO". So we wonder if the transcription factor
hif-1 inactivates "FOXO" somehow, as does PAK1, and this natural product (PPD)
suppresses effectively the growth of both NF1 and NF2 tumors in vivo, as anti-PAK1
products such as Bio 30. Like Bio 30, PPD has no effect on the normal cell growth.

To be continued.

J Int Med Res. 2009 May-Jun;37(3):631-40.

Polyphyllin D Exerts Potent Anti-tumour Effects on Lewis Cancer Cells Under
Hypoxic Conditions.

Ma DD, Lu HX, Xu LS, Xiao W.

Department of Respiratory Medicine, Qilu Hospital, Shandong University,
Jinan, China; Department of Oral Biology, University of Missouri-Kansas
City, Kansas City, Kansas, USA.

Paris polyphylla has been used to treat cancer in China for many years and
components of the plant, such as polyphyllin D, may have potent antiproliferative
effects in vitro. To investigate the potential antitumour effects of polyphyllin
D on cancer cells under hypoxia, Lewis lung cancer cells and mouse tracheal
epithelial cells were cultured with or without polyphyllin D under normoxic
and hypoxic conditions. Proliferation and apoptosis of cells were assayed.
Real-time reverse transcription-polymerase chain reaction was used to quantify
the expression of hypoxia-inducible factor 1 alpha (HIF-1alpha) and vascular
endothelial growth factor (VEGF) mRNA.

Polyphyllin D decreased cell proliferation, increased apoptosis and inhibited
expression of HIF-1alpha and VEGF mRNAs in Lewis cells. These effects were
greater under hypoxic than normoxic conditions. Polyphyllin D did not show
a cytotoxic effect in non-tumour cells (mouse skin fibroblasts and tracheal
epithelial cells). These results suggest that polyphyllin D potentially has
anticancer effects in vitro under hypoxia.



PLoS One. 2009 Jul 27;4(7):e6348.

The HIF-1 hypoxia-inducible factor modulates lifespan in C. elegans.

Zhang Y, Shao Z, Zhai Z, Shen C, Powell-Coffman JA.

Department of Genetics, Development, and Cell Biology, Iowa State University,
Ames, IA, USA.

During normal development or during disease, animal cells experience hypoxic
(low oxygen) conditions, and the hypoxia-inducible factor (HIF) transcription
factors implement most of the critical changes in gene expression that enable
animals to adapt to this stress. Here, we examine the roles of HIF-1 in
post-mitotic aging. We examined the effects of HIF-1 over-expression and
of hif-1 loss-of-function mutations on longevity in C. elegans, a powerful
genetic system in which adult somatic cells are post-mitotic.

We constructed transgenic lines that expressed varying levels of HIF-1 protein
and discovered a positive correlation between HIF-1 expression levels and
lifespan. The data further showed that HIF-1 acted in parallel to the SKN-1/NRF
and DAF-16/FOXO transcription factors to promote longevity. HIF-1 over-expression
also conferred increased resistance to heat and oxidative stress.

We isolated and characterized additional hif-1 mutations, and we found that
each of 3 loss-of-function mutations conferred increased longevity in normal
lab culture conditions, but, unlike HIF-1 over-expression, a hif-1 deletion
mutation did not extend the lifespan of daf-16 or skn-1 mutants.


We conclude that HIF-1 over-expression and hif-1 loss-of-function mutations
promote longevity by different pathways. These data establish HIF-1 as one of
the key stress-responsive transcription factors that modulate longevity in C.
elegans and advance our understanding of the regulatory networks that link
oxygen homeostasis and aging.

2009年8月3日月曜日

Murray Rose: Our OZ "Vegetarian" Hero in Swimming

Murray Rose was born on 6 January 1939 in Scotland, but moved to Australia
with his family at an early age after World War II. He took up swimming
as a boy and was an Olympic Games champion at age seventeen.

Rose became an Olympian for the first time at the 1956 Summer Olympics in
Melbourne. He won the 400 meter and 1500 meter freestyle races and was a
member of the winning team in the 4x200 meter relay. Winning three gold
medals in his home country immediately made him a national hero. He was
the youngest Olympian to be awarded three gold medals in one Olympic Games.

Afterwards, Rose moved to the United States to study at University of Southern
California (USC). At the 1960 Summer Olympics in Rome, Rose again won
an Olympic gold medal. He also won a silver and a bronze bringing his haul
to six medals. Rose continued to compete through his graduation from USC
in 1962. He eventually set fifteen world records. Rose continued to compete
as a masters swimmer.

During his career, he was also known for his strict "Vegetarianism". This
earned him the nickname "The Seaweed Streak". On January 6, 1959, he appeared
as a guest challenger on the TV panel show To Tell The Truth. He is the
patron of the Australian charity 'The Rainbow Club' which teaches disabled
children how to swim. (from: http://en.wikipedia.org/wiki/Murray_Rose)

2009年8月2日日曜日

なぜ極東には偉大な女性政治家が出現しないのか?

東南アジア諸国では、インドのインデラ・ガンジー首相、フィリピンのコラゾン・
アキノ大統領、パキスタンのベナジル・ブット首相、ビルマのアンサン・スーチー
など、歴代の「祖国独立の父」の娘が、祖国の民主主義の危機に際して、立ち上
がって、新しい政権を樹立した(あるいは「独裁政権」に挑戦しつつある)。と
ころが、日本、中国、南北朝鮮、台湾、モンゴルを含む極東地域では、女性が一
度も国の政治の先頭に立ったことがない。

学閥と「OB」(「OG」ではない!) により支えられている永年の封建的な
「官僚主義」がその最大の障害(要因)になっている。

今月の総選挙で、念願の「政権交代」を勝え、民主党政権の下で、ぜひ官僚主義
を打破しようではないか!

2009年7月27日月曜日

メタボ、肥満症はPAK遮断剤であるプロポリスや納豆などで
治療可能かもしれない!

炎症現象には一般に、「PAK」という発癌キナーゼが必須であることが米国で
最近明らかになった。従って、「CD8T細胞」というリンパ球の浸潤を抑える
PAK遮断剤、例えば「Bio 30」などのプロポリスや、ビタミンK2(や
はり、PAK遮断剤)を豊富に含む納豆などで、いわゆる「メタボ」や肥満症を
治療しうる可能性が出てきた。詳しくは、下記を参照されたし。


メタボの原因は脂肪の炎症 薬で治療できるかも?

朝日新聞 (2009年7月27日) より

 メタボリック症候群は、肥満でたまった内臓脂肪に免疫細胞が集まって炎症状
態を起こすことが原因であると、東京大の研究グループがマウスの実験で確かめ
た。糖尿病などの生活習慣病をまねくメタボを免疫を調整する薬で抑えられる可
能性を示した成果で、26日付の米医学誌ネイチャーメディシン電子版に発表し
た。

 東京大の真鍋一郎特任准教授と西村智特任助教ら循環器内科のグループは、蛍
光色素を注射して内臓の脂肪組織の細胞をそのまま観察できる方法を開発、高脂
肪のエサを与えたマウスの内臓の状態を調べた。

 すると、マウスが太って脂肪細胞が大きくなるにつれ、病原体を攻撃する「C
D8T細胞」というリンパ球の一種が出現し、さらに各種の免疫細胞が集まり、
炎症状態になっていることが確認できた。

 CD8T細胞を働かなくしたり、なくしたりしたマウスでは、高脂肪のエサを
与えても、内臓脂肪に免疫細胞が集まる炎症の状態は起こらなかった。さらに、
CD8T細胞がないマウスに、この細胞を入れると、脂肪組織に炎症が起きた。

 また、脂肪組織の炎症が起きているマウスに、この細胞への抗体を与えると、
インスリンが効きやすくなり、血糖値が下がった。

 肥満した人の脂肪組織でも同じようなことが起きているのかはまだ不明だが、
グループの永井良三・東京大教授(循環器内科)は「CD8T細胞を抑える薬な
どでメタボリック症候群による生活習慣病を治療できる可能性が出てきた」とし
ている。(本多昭彦)

(注):
実は、パリ大学医学部の研究グループが2008年に、同様な結果を既に発表して
いる(下記の英文要旨を参照されたし)。

Gastroenterology. 34:1459-69.

Obesity-induced lymphocyte hyperresponsiveness to chemokines: a new mechanism
of Fatty liver inflammation in obese mice.

Bigorgne AE, Bouchet-Delbos L, Naveau S, Dagher I, Prevot S, Durand-Gasselin
I, Couderc J, Valet P, Emilie D, Perlemuter G. (2008)


INSERM, U764, Clamart, France; Univ Paris-Sud, Faculte de Medecine Paris-Sud,
Institut Federatif de Recherche 13, Clamart, France.

BACKGROUND & AIMS: Hepatic lipid retention (steatosis) predisposes hepatitis.
We investigated the mechanisms of lymphocyte homing to fatty liver and
the role of lipopolysaccharide (LPS) in the onset of inflammation in ob/ob
mice.

METHODS: We decreased intestinal bacterial compounds by oral antibiotic
treatment to test the role of endogenous LPS in liver inflammation. Adoptive
transfer of lymphocytes was used to study the respective contributions of
steatosis and lymphocytes to liver inflammation. We tested lymphocyte response
to chemokines by in vitro chemotaxis assays in ob/ob, their lean controls,
and "non-obese ob/ob" mice, generated by controlling caloric intake to distinguish
between the effects of obesity and leptin deficiency.

RESULTS: Antibiotic treatment decreased liver infiltration with CD4(+) T,
CD8(+) T, natural killer (NK)T, B, and NK cells. Adoptive transfer of lymphocytes
from ob/ob or control mice showed that (1) steatosis increased lymphocyte
recruitment to the liver; (2) CD4(+) T, CD8(+) T, and B cells from ob/ob
mice had a greater propensity to migrate specifically to the liver. This
migration was enhanced by LPS. These results were also observed in a model
of high-fat diet-induced obesity. CD4(+) T and B cells were hyperresponsive
to CXCL12 and CXCL13, respectively. Weight normalization in "non-obese ob/ob"
mice decreased liver inflammation, lymphocyte response to chemokines, and
homing to the liver.

CONCLUSIONS: Our study provides the first evidence that liver inflammation
in mice with genetic or diet-induced obesity results from both steatosis
and lymphocyte hyperresponsiveness to chemokines expressed in the liver.
These abnormalities are reversible with weight normalization.

2009年7月20日月曜日

Curcumin could cure both NF2 and TSC (tuberous sclerosis)
by "double" punches, knocking out PAK1 and TOR

This year a group at LSU in Louisiana found that Curcumin blocks the Raptor-TOR
(target of rapamycin) interaction which is essential for the activation
of TOR. The kinase TOR is abnormally activated when the tumor suppressors, TSC1 or
TSC2, dysfunction. The rare but formidable tumors associated with TSC (tuberous
sclerosis), where TSC1 or TSC2 dysfunctions, require TOR for their growth.
In animal experiments anti-TOR drugs such as rapamycin could suppress the
growth of TSC tumors, but none of these anti-TOR drugs is available on the
market as yet. However, the natural anti-cancer product "Curcumin" could
be potentially useful for the treatment of TSC, if its bioavailablity is
dramatically improved by either liposome, CD or propolis in the future, as
mentioned before.

Furthermore, two US groups, at Memorial Sloan-Kettering Cancer Center in
New York and MGH in Boston, reported recently that dysfunction of Merlin,
the NF2 gene product, which is the cause of NF2 tumors, also leads to the abnormal
activation of TOR. We found several years ago that Merlin is a direct PAK1
inhibitor (Hirokawa, Y. et al, 2004). It still remains to be clarified whether
the abnormal activation of TOR is through the kinase PAK1, which is abnormally
activated by the dysfunction of Merlin. Nevertheless, this finding suggests
that "Curcumin" could suppress the growth of NF2 tumors by at least two
ways, inactivating both PAK1 and TOR.

In this context, it would be of great interest to note that the tumor suppressor
"FOXO"/Daf-16 in a tiny nematode called C. elegans blocks the expression
of "Raptor", thereby inactivating "TOR". This potentially interesting finding
was reported in 2004 by Don Riddle's group at University of Missouri. Furthermore,
we recently found that the kinase PAK1 inactivates "FOXO" in this worm.

Thus, natural anti-PAK1 products such as Bio 30 and "Curcumin" would inactivate
"TOR" by reactivating "FOXO" at least in this worm. It would be certainly
worth testing whether this remains true with mammals, too.

To be continued

Beevers CS, Chen L, Liu L, Luo Y, Webster NJ, Huang S. (2009).
Curcumin disrupts the Mammalian target of rapamycin-raptor complex.
Cancer Res. 69, 1000-8.


Department of Biochemistry and Molecular Biology, Feist-Weiller Cancer Center,
Louisiana State University Health Sciences Center, Shreveport, Louisiana
71130-3932, USA.

Curcumin (diferuloylmethane), a polyphenol natural product of the plant
Curcuma longa, is undergoing early clinical trials as a novel anticancer
agent. However, the anticancer mechanism of curcumin remains to be elucidated.
Recently, we have shown that curcumin inhibits phosphorylation of p70 S6
kinase 1 (S6K1) and eukaryotic initiation factor 4E (eIF4E) binding protein
1 (4E-BP1), two downstream effector molecules of the mammalian target of
rapamycin complex 1 (mTORC1) in numerous cancer cell lines. This study was
designed to elucidate the underlying mechanism. We observed that curcumin
inhibited mTORC1 signaling not by inhibition of the upstream kinases, such
as insulin-like growth factor 1 receptor (IGF-IR) and phosphoinositide-dependent
kinase 1 (PDK1). Further, we found that curcumin inhibited mTORC1 signaling
independently of protein phosphatase 2A (PP2A) or AMP-activated protein kinase
AMPK-tuberous sclerosis complex (TSC). This is evidenced by the findings
that curcumin was able to inhibit phosphorylation of S6K1 and 4E-BP1 in
the cells pretreated with PP2A inhibitor (okadaic acid) or AMPK inhibitor
(compound C), or in the cells expressing dominant-negative (dn) PP2A, shRNA
to PP2A-A subunit, or dn-AMPKalpha. Curcumin did not alter the TSC1/2 interaction.
Knockout of TSC2 did not affect curcumin inhibition of mTOR signaling.
Finally, we identified that

curcumin was able to dissociate raptor from mTOR, leading to inhibition
of mTORC1 activity. Therefore, our data indicate that curcumin may represent
a new class of mTOR inhibitor.


Lopez-Lago MA, Okada T, Murillo MM, Socci N, Giancotti FG. (2009)
Loss of the tumor suppressor gene NF2, encoding merlin, constitutively activates
integrin-dependent mTORC1 signaling.
Mol Cell Biol. 29, 4235-49.


Cell Biology Program, Sloan-Kettering Institute for Cancer Research, Memorial
Sloan-Kettering Cancer Center, 1275 York Ave., New York, NY 10065, USA.

Integrin signaling promotes, through PAK1, phosphorylation and inactivation
of the tumor suppressor merlin, thus removing a block to mitogenesis in
normal cells. However, the biochemical function of merlin and the effector
pathways critical for the pathogenesis of malignant mesothelioma and other
NF2-related malignancies are not known. We report that integrin-specific
signaling promotes activation of mTORC1 and cap-dependent mRNA translation.

Depletion of merlin rescues mTORC1 signaling in cells deprived of anchorage
to a permissive extracellular matrix, suggesting that integrin signaling
controls mTORC1 through inactivation of merlin.

This signaling pathway controls translation of the cyclin D1 mRNA and, thereby,
cell cycle progression. In addition, it promotes cell survival. Analysis
of a panel of malignant mesothelioma cell lines reveals a strong correlation
between loss of merlin and activation of mTORC1.

Merlin-negative lines are sensitive to the growth-inhibitory effect of rapamycin,
and the expression of recombinant merlin renders them partially resistant
to rapamycin. Conversely, depletion of merlin restores rapamycin sensitivity
in merlin-positive lines. These results indicate that integrin-mediated
adhesion promotes mTORC1 signaling through the inactivation of merlin. Furthermore,
they reveal that merlin-negative mesotheliomas display unregulated mTORC1
signaling and are sensitive to rapamycin, thus providing a preclinical rationale
for prospective, biomarker-driven clinical studies of mTORC1 inhibitors
in these tumors.

2009年7月19日日曜日

"Combo" Therapy of PAK1-dependent Cancers and NF
by Two Inexpensive Natural Products, Bio 30 and Curcumin

NF (neurofibromatosis) tumors and 70% of human cancers require the oncogenic
kinase PAK1 for their growth. We have recently shown that Bio 30, the CAPE
(caffeic acid phenethyl ester) -rich extract of NZ (New Zealand) propolis
almost completely suppresses the growth of NF tumors, glioma, pancreatic
and breast cancer xenografts in mice. Another natural product called curcumin
was also known to block this kinase somehow. This summer Feng Li's group
at China Medical School reported that curcumin inactivates PAK1, at least
in part by down-regulating the Tyr-kinase ErbB2/HER2 (for detail, see the
abstract below). Several years ago we have shown that both ErbB1 (EGF receptor)
and ErbB2 are required for PAK1 activation (Hong He et al, 2001).

The major problem of curcumin alone for clinical application is its poor
bioavailability (mainly due to its water-insolubility) as previously discussed.
In the future, liposome or CD (cyclodextrin) could be used to solubilize
curcumin for clinical application. Interestingly, CAPE alone shares the same
problem. However, if CAPE is taken with several other anti-cancer polyphenols
in CAPE-based propolis extracts such as Bio 30, both its bioavailability
and anti-cancer potential are dramatically (over 600 times) improved (Maria
Demestre et al, 2009). Considering this finding, it would be worth to combine
curcumin and Bio 30 not only for improving the bioavailability of curcumin,
but also for creating its potential synergy with CAPE and several other
anti-cancer ingredients such as pinocembrin (PIN) in Bio 30.


Cancer Biol Ther. 2009 Jul 13;8(14).

Curcumin suppresses proliferation and invasion in human gastric cancer cells
by downregulation of PAK1 activity and cyclin D1 expression.

Cai XZ, Wang J, Li XD, Wang GL, Liu FN, Cheng MS, Li F.

Department of Cell Biology, Key Laboratory of Cell Biology of Ministry of
Public Health of China, China Medical University, Shenyang, China.

Curcumin (diferuloylmethane), is a natural chemopreventive agent known to
inhibit the proliferation of several cancer cell lines. It has been previously
demonstrated that curcumin is a potent inhibitor of EGF-receptor (EGFR)
tyrosine kinase, but its inhibitive effect on PAK1, a downstream protein
of EGFR, has not been defined. In this paper

we found that curcumin repressed the expression of HER2 and inhibited the
kinase activity of PAK1 without affecting its expression. Silencing HER2
in gastric cancer cells showed that even if PAK1 activity was transiently
strengthened by EGF, curcumin still had a strong inhibitive effect. It should
be emphasized that kinase assay in vitro showed that curcumin could act as
an ATP-competitive inhibitor, which was supported by computer-aided molecular
modeling. Curcumin also downregulated the mRNA and the protein expression
of cyclin D1 and suppressed transition of the cells from G(1) to S phase.
Therefore, curcumin inhibited the proliferation and invasion of gastric
cancer cells. Overall, these results provided novel insights into the mechanisms
of curcumin inhibition of gastric cancer cell growth and potential therapeutic
strategies for gastric cancer.

2009年7月13日月曜日

OSU-03012, the "Synthetic" Anti-PAK1 Drug,
Blocks the Growth of NF2 Tumor Xenografts in Mice.

In 2007 a group led by Matthew Ringel at OSU (Ohio State University) found
that a synthetic compound called OSU-03012 selectively inhibits PAK1 in
vitro with IC50 around 1 micro M, and blocks the growth of several thyroid
cancer cell lines. In other words thyroid cancer is among PAK1-depenent
cancers. This compound is a derivative of the cyclooxygenese-2 inhibitor
"celecoxib", and was initially developed as a PDK1 inhibitor by Ching-Shih Chen's
group at OSU in 2004. It eventually inactivates another kinase AKT
by inhibiting the kinase PDK1. However, since it inhibits the growth of
PAK1-dependent (and AKT-independent) cancer cell lines such as PC-3 (PTEN-deficient
prostate cancer) and pancreatic cancer cells, it had been suggested that
OSU-03012 might block the kinase PAK1. In 2009 OSU-03012 (200 mg/kg, daily)
was found by Long-Sheng Chang's group at OSU to suppress the growth of NF2-deficient tumors
(Schwannoma) xenografts in mice by more than 50% (for detail, see the abstract below).

It should be worth to point it out that like ARC (Artepillin C) from Brazilian
green propolis, OSU-03012 is among the first single chemical drugs that
were shown to block the growth of NF2 tumors in vivo, although Bio 30
(100 mg/kg, twice a week) a mixture of several anti-cancer ingredients
including CAPE, was proven to block completely the growth of NF2 tumors in vivo, and is inexpensively available
on the market. If OSU-03012 is successful in the time and money-consuming
clinical trials for these PAK1-dependent cancers and NF tumors, this "synthetic"
compound would become available on the market in a decade or so... Hope
(unlike FK228) this drug can pass BBB (blood brain barrier) so that it works
on formidable brain tumors such as glioma and NF2 tumors.

Eur J Cancer. 2009 Jun; 45(9):1709-20

Growth inhibitory and anti-tumour activities of OSU-03012, a novel PDK-1
inhibitor, on vestibular schwannoma and malignant schwannoma cells.

Lee TX, Packer MD, Huang J, Akhmametyeva EM, Kulp SK, Chen CS, Giovannini
M, Jacob A, Welling DB, Chang LS.

Department of Otolaryngology, The Ohio State University College of Medicine,
Center for Childhood Cancer, The Research Institute at Nationwide Children's
Hospital, Columbus, OH, USA.

BACKGROUND: Vestibular schwannomas (VS) frequently express high levels of
activated AKT. Small-molecule inhibitors of AKT signalling may have therapeutic
potential in suppressing the growth of benign VS and malignant schwannomas.
METHOD: Primary VS and Schwann cells, human malignant schwannoma HMS-97
cells and mouse Nf2(-/-) Schwann cells and schwannoma cells were prepared
to investigate the growth inhibitory and anti-tumour activities of OSU-03012,
a celecoxib-derived small-molecule inhibitor of phosphoinositide-dependent
kinase-1. Cell proliferation assays, apoptosis, Western blot, in vivo xenograft
analysis using SCID mice and immunohistochemistry were performed.

RESULTS: OSU-03012 inhibited cell proliferation more effectively in both
VS and HMS-97 cells than in normal human Schwann cells. The IC5) of OSU-03012
at 48h was approximately 3 micro M for VS cells and HMS-97 cells, compared
with the IC(50) of greater than 12 micro M for human (normal) Schwann cells.
Similarly, mouse Nf2(-/-) schwannoma and Nf2(-/-) Schwann cells were more
sensitive to growth inhibition by OSU-03012 than wild-type mouse Schwann
cells and mouse schwannoma cells established from transgenic mice carrying
the NF2 promoter-driven SV40 T-antigen gene. Like VS cells, malignant schwannoma
HMS-97 cells expressed high levels of activated AKT. OSU-03012 induced apoptosis
in both VS and HMS-97 cells and caused a marked reduction of AKT phosphorylation
at both the Ser-308 and Thr-473 sites in a dose-dependent manner.

In vivo xenograft analysis showed that OSU-03012 was well tolerated and
inhibited the growth of HMS-97 schwannoma xenografts by 55% after 9 weeks
of oral treatment. The anti-tumour activity correlated with reduced AKT
phosphorylation.

CONCLUSION: OSU-03012 is a potential chemotherapeutic agent for VS and malignant
schwannomas.

2009年7月7日火曜日

PAK1-LC8 (Dynein Light Chain) Interaction
Required for Nuclear Import of PAK1.

PAK1 is an oncogenic Ser/Thr kinase which is activated by several cytoplasmic
proteins such as GTPases (Rac and CDC42), SH3 adaptor proteins (PIX and
NCK), and Tyr-kinase (ETK). This kinase is required for the growth of more
than 70% of human cancers, including breast and prostate cancers, as well
as formidable brain tumors such as glioma and NF (neurofibromatosis) tumors.

Recently John Williams' group at Thomas Jefferson University in Philadelphia
found that Dynein Light Chain (LC8) also binds residues 212-222 of PAK1
which is adjacent to the essential NLS (nuclear localization site, residues
243-245), and their interaction leads to the dimerization of PAK1 through
NLS and is required for the EGF-induced nuclear import of the cytoplasmic
PAK1 in MCF-7, an estrogen-dependent breast cancer cell line. Since PAK1
phosphorylates the nuclear estrogen receptor (ER) at Ser 305 for the ER
activation which leads to the estrogen-dependent malignant growth of this
cell line, LC8 appears to contribute to the oncogenicity of PAK1 by nuclear
import.


PLoS One. 2009 Jun 26;4(6):e6025

Interaction with LC8 is required for Pak1 nuclear import and is indispensable
for zebrafish development.

Lightcap CM, Kari G, Arias-Romero LE, Chernoff J, Rodeck U, Williams, John
C.

Department of Biochemistry and Molecular Biology, Thomas Jefferson University,
Philadelphia, PA, USA.

Pak1 is a serine/threonine kinase implicated in regulation of cell motility
and survival and in malignant transformation of mammary epithelial cells.
In addition, the dynein light chain, LC8, has been described to cooperate
with Pak1 in malignant transformation of breast cancer cells. Pak1 itself
may aid breast cancer development by phosphorylating nuclear proteins, including
estrogen receptor alpha. Recently, we showed that the LC8 binding site on
Pak1 is adjacent to the nuclear localization sequence (NLS) required for
Pak1 nuclear import.

Here, we demonstrate that the LC8-Pak1 interaction is necessary for epidermal
growth factor (EGF)-induced nuclear import of Pak1 in MCF-7 cells, and that
this event is contingent upon LC8-mediated Pak1 dimerization. In contrast,
Pak2, which lacks an LC8 binding site but contains a nuclear localization
sequence identical to that in Pak1, remains cytoplasmic upon EGF stimulation
of MCF-7 cells.

Furthermore, we show that severe developmental defects in zebrafish embryos
caused by morpholino injections targeting Pak are partially rescued by co-injection
of wild-type human Pak1, but not by co-injection of mutant Pak1 mRNA disrupting
either the LC8 binding or the NLS site.

Collectively, these results suggest that LC8 facilitates nuclear import
of Pak1 and that this function is indispensable during vertebrate development.

2009年6月19日金曜日

「シクロデキストリン」の食品や医薬への応用

シクロデキストリンとは、環状オリゴ糖で、分子の外側は親水性で、内側は疎水
性である。従って、疎水性の低分子物質を、その分子の内側に取り込み、水溶化
しうる働きを持つ。

詳しくは、ブドウ糖が連なってできたオリゴ糖の両端が繋がり輪の形になってお
り、構造的には、フタと底のないカップのような分子である。そして、この空洞
部分に様々な分子を取り込んだり(包接)、逆に取り込んだ分子を条件に応じて
中からゆっくりと放出させたり(徐放)といった作用を示めす。この性質を応用
して様々な分野でいろいろな商品に利用されている。ちなみに、シクロデキスト
リンは、ジャガイモやトウモロコシ等のデンプンを原材料としている。

具体的な商品化の一例としては、臭い成分を取り込む作用を活かした消臭剤への
活用やカテキンやイソフラボンなどの有効成分の苦み渋みを低減する目的で飲料
への利用、また卵黄・バター等からコレステロールの除去効果を利用した食品分
野への応用などがある。さらに、機能性食品やサプリメントへの応用として、例
えば、コエンザイムQ10など難水溶性のため吸収されにくい機能性成分を、シクロ
デキストリンにより水溶性化させ吸収を高めるといった利用もなされている。

我々が最近興味をもち始めたのは、シクロデキストンをプロポリス、花椒エキス、
クルクミン、イバメクチン、納豆のビタミンK2(メナキノンー7)など難水溶
性の天然制癌(PAK遮断) 物質の水溶化に利用して、その体内への吸収を促進
するばかりではなく、その独特の苦味や匂いを軽減する働きを活用することにあ
る。さらに、シクロデキストンは、その空洞内に低分子だけを選択的に取り込む
ので、蛋白や多糖類などの高分子をも含む酵母や植物エキスから、低分子の抗癌
物質、例えば「ピノセンブリン」だけを選別/抽出しうるツールとしても利用できる
可能性があるからだ。

数年前、シクロケム (社長、寺尾啓二) が岡山理科大学との共同研究で、天然の
アルファー・シクロデキストン(alphaーCD、2%)によるクルクミンの水溶化(最大溶解度、
0。3 mg/ml) に成功したという報告が出ている。 動物実験で、この
CDークルクミンが実際に癌やNF腫瘍の増殖抑制に有効というデータはまだ出
ていないようだが、例えば、担癌マウス (体重20g)にこの溶液を週2回ずつ、
最大0。4 ml (つまり、6 mg/kg)腹腔注射して、有意な効果 (例えば、
50%以上の増殖阻害) が得られれば、この新しいクルクミン製剤による癌治療
の早期実用化が有望となろう。なお、クルクミンの水溶化は、リポソームやベー
タCD誘導体でも最近、米国やインドでそれぞれ成功しているが、これらの可溶
化剤(後者)は天然物ではないので、治療への実用化(各々の政府から市販許可
を得る)には、まだ時間が相当かかるだろう。

参考までに、リポソームによる包接の場合を一例としてあげると、担癌マウスに
クリクミン投与 (20 mg/kg) で、すいぞう癌の増殖が50%以上抑制される
という結果が、テキサスのMDアンダーソン癌センターから、つい最近報告されている。

Anticancer Res. 2009 Jun; 29: 1895-9.

Determination of minimum effective dose and optimal dosing schedule
for liposomal curcumin in a xenograft human pancreatic cancer model.

Mach CM, Mathew L, Mosley SA, Kurzrock R, Smith JA.

Department of Gynecologic Oncology, Division of Surgery, P.O. Box
301439-UNIT 1362, Houston, Texas 77230-1439, U.S.A.
jasmith@mdanderson.org

BACKGROUND: Curcumin is a food chemical present in tumeric (Curcuma longa)
that has pharmacological activity to suppress carcinogenesis and inhibits
multiple signaling pathways such as nuclear factor kappaB (NF-kappaB),
cyclooxygenase-2 (Cox-2) and interleukin-8 (IL-8). Oral curcumin has poor oral
bioavailability limiting its clinical activity;
however, a patent pending liposomal formulation of curcumin was developed
to improve drug delivery and has demonstrated activity in multiple cancers.
This study was designed to determine the minimum effective dose (MED)
as well as the optimal dosing schedule of liposomal curcumin
in a xenograft mouse model of human pancreatic cancer.

MATERIALS AND METHODS: The MED determination and optimal schedule was evaluated
in female athymic nude mice injected subcutaneously with MiaPaCa-2 cells.
Dosing was initiated at an average tumor size of 5mm. For the MED, mice
were treated with the following dose levels of liposomal curcumin: no treatment,
liposome only, 1 mg/kg, 2 mg/kg, 5 mg/kg, 10 mg/kg, 20 mg/kg and 40 mg/kg
given by tail vein injection three times weekly for 28 days. For the optimum
dosing schedule, three additional schedules were evaluated and compared
to the control of three times weekly; daily (five days per week), every
four days, and weekly for 28 days. All mice were weighed and tumor measurements
taken three times weekly to evaluate toxicity and efficacy.

RESULTS: The 20 mg/kg dose had the greatest decrease in tumor growth at
52% decrease in tumor growth when compared to no treatment control mice.
MED was determined to be 20 mg/kg and was used for the optimal dosing schedule
determination. Daily dosing and three times per week dosing had greater
inhibition of tumor growth with no discernable difference than once weekly
or every 4 day dosing. No toxicity was observed at any dose or schedule.

CONCLUSION: The MED for liposomal curcumin is 20 mg/kg given once daily
three times per week to achieve optimal tumor growth inhibition. This was
dose recommended for additional preclinical studies to define safety and
tolerability of liposomal curcumin in rat and dog models.

続く

2009年6月18日木曜日

"PIN-MITE": a new Pinocembrin (PIN)-rich health care product

Pinocembrin (PIN) content in Bio 30 is 11% (110 mg/g, dry weight), the highest
among propolis samples around the world. PIN is a flavonoid rich in pine
trees. A cluster of genes such as CHS and CHI for its biosynthesis was cloned
from red pine several years ago, and now it becomes possible to mass produce
PIN in E. coli or yeasts (S. cerevisiae or S. pombe), using glucose and
Phe as two major nutrient sources. We are planning to mass produce PIN in
yeast under the control of Hsp16 (heat-shock) gene promoter by heat shock
treatment, to make a new PIN-rich health care product called "PIN-Mite"
(anti-cancer/NF remedy or "elixir") as an alternative of Bio 30 or "Natto".


Why are we so interested in PIN? it has been known for some years that
like CAPE (caffeic acid phenethyl ester) , PIN is anti-inflammatory, suggesting
the possibility that it might block the kinase PAK1 which is essential for
inflammation, in particular calcium-release from mast cells. This year it
was reported by a Japanese group that like CAPE and ARC (artepillin C),
PIN blocks angiogenesis which also requires PAK1. Since both CAPE and ARC,
the major anti-cancer ingredients in propolis, block PAK1, it is almost
certain that PIN also blocks the oncogenic PAK1 signaling somehow. We are
asking a friend of mine in Japan to confirm this notion directly in cell
culture, by testing the effect of PIN on p-Raf 1 (Ser 338) which is a direct
indicator of PAK1's kinase activity.

However, like CAPE and ARC, PIN is water-insoluble (therefore its bioavailability
would be poor if alone). Lipids in propolis solubilize these water-insoluble
PAK1-blockers. That is a reason why CAPE in propolis such as Bio 30 is far
better (more effective) than CAPE alone in vivo. However, there must be
another reason. Bio 30 is 600 times more effective than CAPE alone in vitro
(cell culture). Bio 30 contains several polyphenols/flavonoids, in addition
to CAPE: among them PIN is the most abundant (110 mg/g), almost 10 times
more than CAPE (only 12 mg/g). Thus, PIN and several other anti-cancer flavonoids
in Bio 30 must work synergestically with CAPE to suppress the growth of
PAK1-dependent cancers/NF.

An OZ scientist developed a unique salty yeast extract called "Vegemite"
as a spread in 1922. It has been sold by a major food company called "Kraft"
. "Vegemite" for OZ school kids is equivalent to "peanuts butter" for US
counterparts. They are the most favourite spreads to make their lunch sandwiches
tasty. Well, can we make a hybrid of "Vegemite and PIN-rich yeast extract,
which we might call "PIN-MITE"? YES, WE CAN! Alternatively, can we make
another hybrid product of "Natto" and PIN-rich yeast extract? YES, WE CAN!
As previously mentioned, Natto is a traditional Japanese fermented sticky
soy-bean product, and the richest source of vitamin K2, in particular menaquinone-
7 (MK-7). Like PIN, MK-7 is most likely to block PAK1, and has been shown
to suppress the growth of PAK1-dependent cancers such as pancreatic and ovarian
cancers.

Since PAK1 shortens the life span, anti-PAK1 products in propolis such as
CAPE, ARC and PIN as well as MK-7 in Natto would prolong our life in good
health, protecting us from cancers, NF, inflammation such as asthma and
arthritis, and aging diseases such as AD (Alzheimer's).

Pearl S. Buck (1892-1973), the author of "Good Earth" and the 1938 Nobel
laureate in Literature for bridging the West and East cultures, once said:
a hybrid child is far better than each of his or her (more pure) parents,
because two different (complementary) set of genes cause a synergy. Barack
Obama has proved it at the 2008 US election. Better than the more Anglo-Saxon
(or WASP) candidates Hillary Clinton and John McCaine! Like PSB, Obama
understands both sides: East and West, Christian and Moslem, Poor and Rich,
Conservative and Liberal, Republican and Democrat, Old and Young. His vast
knowledge and tolerance is the major power for governing US and the rest
of our world. So his handsome picture (or PSB's young portrait) could be
used for the label of our new mighty spread "PIN-Mite" jars or bottles for
the world-wide promotion...

To be continued.

2009年6月10日水曜日

Opening a "PRC" (Propolis Research Center)?

Chemical composition of propolis varies from one sample to another, because
honey bees in different areas such as NZ (New Zealand) and Brazil collect
entirely different things from local flowers and trees to make their beehives.
For instance, regarding the major anti-cancer ingredients, NZ propolis
is rich in CAPE (caffeic acid phenethyl ester), while Brazilian green propolis
is rich in ARC (Artepillin C). However, all these propolis samples share
a common biological property. They are anti-bacterial, anti-viral, stimulate
immune system, and block the oncogenic kinase PAK1 which is not only essential
for the growth of more than 70% of human cancers including breast and prostate
cancers as well as NF (neurofibromatosis) tumors, but also shortens our
life span.

Thus, we are potentially interested in opening a unique propolis research
center (PRC) to develop the most potent propolis, which block PAK1 most
efficiently, by our own hands. We have recently developed an inexpensive
and quick in vivo bioassay system to quantify the anti-PAK1 (so-called "elixir
of life") activity of natural products such as propolis. The special strain
(CL2070) of a tiny transparent nematode called C. elegans contains an extra
chimeric gene called Hsp16:GFP which expresses GFP (green fluorescent protein)
under the control of the promoter of Hsp16 gene. GFP is derived from a
fluorescent jellyfish, and glows green when it is exposed to blue light.
This GFP technology was developed by the three 2008 Nobel laureates in Chemistry,
Osamu Shimomura, Martin Chalfie and Roger Tsien. Hsp16 gene produces a
heat shock protein which contributes to the prolonged life span. and heat
endurance (thermal resistance). This gene requires a transcription factor
called "FOXO" which is normally suppressed by the kinase PAK1. So when PAK1
in this worm is blocked by the treatment with propolis, FOXO would be highly
activated and consequently the promoter of Hsp16 gene is activated after
a brief heat-shock to induce the GFP production in this worm. The stronger
its anti-PAK1 activity, the brighter the worm glows.

Using this GFP nematode system, we could screen for the most potent propolis
sample. In both mouse and this worm systems, we have shown that Bio 30,
CAPE-based NZ propolis extract, is much more potent than GPE, ARC-based
Brazilian green propolis extract. In the future we would like to develop
a new propolis which is even far more potent than Bio 30, hopefully. For
this specific purpose, we are planning to open a PRC of our own, a new bee
farm where we grow both honey bees and several selected flowers to harvest
a unique propolis of our own, and determine which combination of flowers
would be the best source for the production of the most potent propolis.
To initiate such a pilot PRC, I shall start a bee farm at my own home garden,
which was recently bought in Brunswick West, outskirts of Melbourne, hopefully
in the near future, following the instruction of an e-book "Bee Keeping
for Beginners" or a paperback "The Backyard Beekeeper: An Absolute Beginner's Guide
to Keeping Bees in Your Yard and Garden"...

To be continued

2009年6月8日月曜日

A New Challenge: Bee Farming at Home

Last Saturday (June 6th), I finally managed to buy an old brick house (built
in an early 1940s) standing on a small land of around 400 m2, Brunswick
West in outskirts of Melbourne. It has two bedrooms, a separate extra bed/office
room (to be used as a library for me), and a relatively large garden space.
Around October 2, I shall move in this new home from my humble one-room
cabin (called "Abe's Cabin") in North Melbourne which I rent just for 12
months. Abe is a nick name of Abraham Lincoln who spent his childhood in
a log cabin, before he became a lawyer and eventually the 16th US President
to abolish the slavery for good.

My very first task at this home is to start growing a few aromatic flower
plants such as Brunflesia and Jasmin in this garden. Then I shall start
trying bee farming at home, hoping to produce a home-made "aromatic" propolis
and honey.

As I mentioned before, propolis is the major natural source of potent anti-cancer
ingredients such as CAPE (caffeic acid phenethyl ester) and ARC (Artepillin
C) that block selectively the oncogenic kinase PAK1. This kinase is required
for the growth of more than 70% of human cancers and NF (neurofibromatosis)
tumors as well as several aging diseases such as Alzheimer's (AD) and arthritis.

In addition propolis is good for protecting us from infectious (bacterial/viral) diseases
such as AIDS and chicken/swine flu. Since this kinase normally shortens
our life span by inactivating the tumor suppressor "FOXO", anti-PAK1 products
such as propolis would prolong significantly our life in good health.

I hope my new neighbors in this suburbs would not reject this bee farming
project of mine, because it would eventually improve their QOL (quality
of life) as well...

to be continued