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Rapid replication is responsible for introducing genetic defects causing diseases. When an additional piece of nucleotide is attached to a DNA string, it is called insertion or a piece of DNA is removed from the DNA string; it is called deletion or structural inversion of DNA is responsible for mutations.

Since the gene in a DNA codes for proteins, insertion and deletion on DNA have catastrophic effects on protein synthesis.

In glioblastomas, three major changes occur on chromosomes (C-7, C-9, and C-10) and two minor changes occur on chromosomes (C-1 and C-19). These mutations are responsible for causing brain cancers in humans. In a normal human cell, chromosome-7 which is made of 171 million nucleotide base pairs and carries 1378 genes.

When insertion occurs on chromosome-7, 97% of glioblastoma patients are affected by this mutation. On the other hand, a different mutation occurs on chromosome-9 which is made of 145 million nucleotide base pairs and it carries 1076 genes.

A major deletion of a piece of DNA occurs on chromosome-9, which results in 83% patients who are affected by this mutation. A minor deletion of DNA also occurs on chromosome-10 which is made of 144 million base pairs and it carries 923 genes.

Although it is a minor deletion of a piece of DNA, it contributes to 91% patients with glioblastoma. To a lesser extent, small mutation occurs on

chromosome-1

(the largest chromosome in our genome). It is made of 263 million nucleotide base pairs and carries 2610 genes), and chromosome-19 (it is made of 67 million base pairs and carries 1592 genes) is also implicated in some forms of glioblastomas. All known glioblastomas causing genes are located on five different chromosomes and carries a total of 9579 genes.

It appears impossible to design drugs to treat glioblastomas since we do not know which nucleotide on which gene and on which chromosome is responsible for causing the disease.

With the completion of 1000 Human Genome Project, it becomes easier. By simply comparing the patient’s chromosomes with the 1000 genomes, letter by letter, word by word, and sentence by sentence, we could identify the difference called the variants with precision and accuracy, the exact variants or mutations responsible for causing the disease.

Once the diagnosis is confirmed, the next step is how to treat the disease.

With the quinone ring, I could introduce different combinations of aziridine rings and carbamate moieties and could create havoc for glioblastomas. My major concern was how toxic this compound would be to the human brain cells. Fortunately, brain cells do not divide, only cancer cells divide. Our rational drug design work began in the University of London, England, and completed in the Laboratory of the National Cancer Institute (NCI), of the National Institutes of Health (NIH), in Bethesda, Maryland, USA.

Over this period, we conducted over 500 experiments which resulted in 200 novel drugs.

They were all tested against the experimental animal tumors.

Forty-five of them were considered valuable enough to be patented by the US Government (US Patent 4,146,622).

Radiolabeled studies showed that AZQ has the ability to cross organ after organ, cross the blood-brain barrier, cross the nuclear membrane, and attack the nuclear DNA shutting off the gene. X-ray studies showed that the radioactivity is concentrated in the tumor region. For the discovery of AZQ, I was honored with the “2004 NIH Scientific Achievement Award,” one of America’s highest awards in Medicine and I was also honored with the India’s National Medal of Honor, “Vaidya Ratna,” a gold medal (see Figures 1–4).

2004 NIH Scientific Achievement Award presented to Dr.

Elias Zerhouni, the director of NIH during the NIH/APAO award ceremony held on December 3, 2004.

Khan is the discoverer of AZQ (US Patent 4,146,622), a novel experimental drug specifically designed to shut off a gene that causes brain cancer for which he receives a 17-year royalty for his invention (license number L-0I9-0I/0).

To this date, more than 300 research papers have been published on AZQ. The award ceremony was broadcast live worldwide by the Voice of America (VOA). Khan is the first Indian to receive one of America’s highest awards in Medicine. Hameed Khan, discoverer of anticancer AZQ, after receiving 2004, Vaidya Ratna, the gold medal, one of India’s highest awards in Medicine at the Rashtrapati Bhavan (Presidential Palace), in Delhi, India, during a reception held on 02 April 2004. Hameed Khan, a scientist at the National Institutes of Health (NIH), USA, an American scientist of Indian origin was awarded on April 2, 2004.

Vaidya Ratna, the gold medal, one of India’s highest awards in Medicine for his discovery of AZQ (US Patent 4,146,622) which is now undergoing clinical trials for treating bran cancer. Coronary artery disease is complex involving about 60 genomic variants (genes).

All variants are not clustered on any specific chromosome.

These variants are dispersed across the entire genome.

Although all variants have not been sequenced, we can shut off only the mutated gene without knowing the sequence of all other genes.

As I mentioned above in the “Cancers” section, the mutated gene grows rapidly forming the tumor.

As it grows, it uses glucose as a source of energy, which is broken down to produce pyruvic acid.

In the presence of acid, the analogs of aziridine and carbamate are activated to generate carbonium ion which attack the tumor DNA shutting off their genes. While we may someday be able to sequence all 60 genes associated with the coronary artery disease, presently, we can single out and identify the mutated gene bound complex using radiolabeled aziridine and carbamate.

The following example explains how some arrhythmias causing genes could be identified and how drug could be designed to shut off these genes.

The term “QT” refers to the segment of an electrocardiogram, which measures the duration of time for the heart to relax after a heartbeat. In long QT syndrome, the duration of time is abnormally prolonged and creates a vulnerability to dangerous arrhythmias [22].

Ever since the syndrome was described in 1957, researchers have engaged in a genetic race to identify the genes associated with long QT syndrome, which currently includes 17 genes. Three genes, KCNQ1 , KCNH2 , and SCN5A , had sufficient evidence to be implicated as “definitive” genetic causes for typical long QT syndrome.

Four other genes had strong or definitive evidence

supporting

their role in causing atypical forms of long QT syndrome, presenting in the newborn symptoms associated with heart block, seizures, or delays in development. Once the mutated genes are identified, we could design drugs to shut off these genes as described in the “Cancers” section.

Alois Alzheimer identified the microscopic changes in the brain of a patient with the memory loss. He was the first physician to identify the disease in a 50-year-old woman who suffered from psychosis and who died within 4 years. Using special dyes, he stained the brain tissues which carried abnormal protein deposit around her brain which controlled brain function.

He identified two kinds of legions of amyloid patches which he mistakenly thought was fatty patches and now turned out to be proteins.

He observed a patch of fatty deposit on the top of the brain cells called plaques and the legions inside the nerve cells called tangles.

He accurately correlated the abnormal protein deposits around brain cells with the controlled of brain function [23, 24, 25, 26]. Today, we know that the age is the single most risk factor for developing Alzheimer. By age 65 or older, the risk for developing Alzheimer is about

10%

, and by age 85 or older, the risk factor is as high as 40 or 50%.

When he performed the autopsy of many senile persons, Dr.

Alzheimer found the same plaques and tangles in many other samples.

Early onset or late onset of Alzheimer resulted in the epidemic of Alzheimer.

When comparing a normal brain with the Alzheimer brain, we find that the Alzheimer brain has shrunken and there is a concentration of plaques and tangles in neurons. In healthy brain cells, we see occasional plaques and tangles. It defines the disease; the plaque and tangles start building up as we grow old, and over years and decades, the symptoms begin to develop. Symptoms include memory loss and decrease in ability of learning and recall. Early onset affects cognition which encompasses memory and other mental functions such as erosion of attention, thinking, reasoning, visual functions, spatial function, and dementia with memory loss and other cognitive functions resulting in mental impairment which affects to the degree interfering with the daily life. Recent studies confirm that Alzheimer is an irreversible brain disorder which slowly destroys memory and thinking skills.

The damage to the brain is not particularly associated to any specific gene, but the presence of the one form of the apolipoprotein E (APOE) is a suspect gene whose presence does increase a patient’s risk for developing Alzheimer. The early onset of Alzheimer is

associated

with three single gene mutations: first, the presence of an amyloid precursor protein (APP) located on chromosome-21; the presence of presenilin 1 (PSEN1) on chromosome-14 and the presence of presenilin 2 (PSEN2) located on chromosome-1. All three chromosomes are very large and carry hundreds of genes. For example, chromosome-1 is the largest chromosome in the genome. It is made of 163 million nucleotide bases carrying 2610 genes. Chromosome-21 is made of 50 million nucleotide bases carrying 337 genes, while chromosome-14 is made of 109 million nucleotide bases carrying 1173 genes.

A recent 7 million Utah population study identified two additional genes RAB10 located on chromosome-2 (which is made of 155 million nucleotide bases and carry 1798 genes) and SAR1A gene located on chromosome-10 (which is made of 144 million nucleotide bases and carry 983 genes) associated with the formation of plaques and tangles.

Mutations on these genes may be

associated

with the onset of Alzheimer. Of all the genes on these chromosomes, only five single-gene mutations are associated with the early onset of the Alzheimer, it is the greatest challenge to design drugs to attack only the mutated genes.

As I said above in the “Cancers” section, the good news is that the only mutated genes grow rapidly using glucose as a source of energy.

It is the acid which activates the aziridine and carbamate moieties producing powerful carbonium ion which attack N-7 guanine of DNA and shut off only the mutated genes. Using C-14 radiolabeled aziridines, we can identify the mutated gene which form the aziridine/protein complex as described in the “Cancers” section. It is well known that using the TFT dye, which is 3,6-dimethyl-2-(4-dimethlaminophenyl)-benzothiazoline, could be used to stain the plaques and tangles of Alzheimer tissues.

Using TFT dye as a carrier for the

aziridine

and carbamate moieties, we could design drugs to attack the mutated DNA to shut off genes which form plaques and tangles to prevent the progress of Alzheimer.

In the above “Cancers” section, I have described in detail how I had used quinone as a carrier for aziridine and carbamate ions in designing AZQ to attack the brain tumor DNA to shut off genes for treating brain cancer. Similarly, the analogs of benzothiazoline dyes could be used to carry aziridine and carbamate moieties to attack the plaque and tangle DNA and shut off genes responsible for causing Alzheimer.

Could I use the same rational drug design and introduce a novel method for treating breast tumor? Although mutations on BRCA1 gene located on chromosome-17 (which is made of 92 million nucleotide bases carrying 1394 genes) have been identified years ago responsible for causing breast cancer, we wonder why it has been so difficult to design drugs on rational basis to treat breast cancer.

By the time the breast cancer diagnosis is confirmed in a patient, the BRCA1 has accumulated more than 3000 mutations. Genotyping of the blood would also show the existence of many cells carrying mutated cells responsible for creating secondary deposits. It is also believed that by the time breast cancer is confirmed, metastatic cancer cells have already been spread from liver lung on its way to brain. Since all other organs including breast and liver could be removed and replaced by organ transplant except brain, I thought that protecting brain is utmost important to save life. Once AZQ is developed to protect the brain, I could focus on the breast and prostate cancers.

Recent, radiolabeled studies showed that male hormone testosterone has great affinity for female organs like breast, ovary, and fallopian tube cells.

On the other hand, estrogen, the female hormone, has great affinity for male prostate gland.

By attaching multiple aziridine rings and carbamate ions to both hormones, I could design novel drugs to attack the breast and the prostate cancer.

Now, I found that I could go even further by attaching more than four aziridine and carbamate moieties to both male and female hormones.

In a breast tumor, within the start and stop codons, BRCA1 gene has captured over 200,000 nucleotide bases.

These mutations are caused by exposure to radiations, chemical or environmental pollutants, viral infection, or genetic inheritance.



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