HTML 5 version of DOI: 10.2174/1871520620666200313163801 | https://anti-cancer.eu
Small typographical inaccuracies are possible. For scientific application of information use the publisher's licensed print and/or on-line versions! 1
THEORETICAL ANALYSIS FOR THE SAFE FORM AND DOSAGE OF AMYGDALIN
PRODUCT
Vasil Tsanov, Hristo Tsanov
ABSTRACT
This article presents a theoretical analysis of the safe dosage form and its dosage of the amygdalin
derivative. By making a precise socio-anthropological analysis of the life of the ancient people of Botra [1]
(Hunza people, Burusho / Brusho people), we come to the hypothesis, which is confirmed by two proofs,
through a number of modern quantum-mechanical, molecular-topological and bio-analytical checks. A
convenient, harmless, form of amygdalin derivative is available that has the same biological and chemical
activity and could be used in conservative clinical oncology. The article also presents a theoretical comparative
analysis of biochemical reactivity in in vivo and in vitro media, by which we also determine the recommended
dosage for patient administration. Based on a comparative analysis of the data, obtained in published clinical
studies of amygdalin, is presented and summarized a scheme of the anti-tumor activity of proposed by the
author’s molecular form.
KEYWORDS
oncology, amygdalin, PM6, PM7, TD-DFT, pharmacological activity
1. Introduction
The apricot is a fruit known to people for millennia. Archaeological excavations in the ancient Armenian
city of Shenchovit near Yerevan revealed overlaid apricot excavations dating back to 6,000 years BC. The
first written mention of apricot was 4,000 years ago in a letter from a Chinese resident. The well-known apricot
comes from a variety of the high-mountainous region of Hindu Kush - Central Asia, where the borders of
China, Tajikistan, Afghanistan and Pakistan meet today. Natural forest and very old apricot trees can still be
found in northeast China and the Caucasus.
HTML 5 version of DOI: 10.2174/1871520620666200313163801 | https://anti-cancer.eu
Small typographical inaccuracies are possible. For scientific application of information use the publisher's licensed print and/or on-line versions! 2
It is a well-known fact by derontologists that the Hunzi people that inhabited the highlands of northern
Pakistan, not far from where the apricot originates, are the healthiest and longest-lived people in the world.
According to researchers and medical scientists who studied the life of the Huns in their natural environment
in the 1950s and 1960s, one hundred percent of them had perfect vision, and cancer, heart attack, high blood
pressure, high cholesterol and even appendicitis and gout were unknown states for them.
Throughout the year, their diet was rich in dried fruits and nuts, with apricots and apricot kernels
predominating, and their main source of fat was apricot seeds. Apricots were an important part of the Hunzi
life.
The apricot kernels contain an average of 21% protein and 52% vegetable oil and are widely used as a
substitute for almonds in the food, cosmetic and pharmaceutical industries. Due to its high content of
amygdalin, apricot seeds are a source of vitamin B17 and are used in alternative medicine for cancer therapy.
The American Cancer Society notes that apricots, as well as other carotene-rich fruits, reduce the risk
of cancer of the larynx, esophagus and lungs.
1.1. Pharmacological activity of amygdalin.
Amygdalin is a nitrile containing a diglycoside compound of the general formula C
20
H
27
NO
11
, molecular
weight 457.42, with the structure D-mandelonitrile-β-D-glucoside-6-β-glucoside [2] and the structural formula
fig.1.
Fig. 1. Structural formula and full chemical name of Amygdalin
Amygdalin is non-toxic, but under the action of digestive juices and enzymes in the blood it releases
HCN, which, even at relatively low concentrations, is even deadly.
Numerous studies have been performed to prove its antitussive [2] and antiasthmatic [3] effects, analgesic
[4÷9], gastro-enterologic [10,11], promoting apoptosis of human renal fibroblast [12], boosting immunity
synthesis [13÷15] (including to increase polyhydroxyalkanoates in induced human peripheral blood T-
lymphocyte proliferation), anti-diabetic properties [16,17] (including inhibiting alloxan in hyperglycemia),
HTML 5 version of DOI: 10.2174/1871520620666200313163801 | https://anti-cancer.eu
Small typographical inaccuracies are possible. For scientific application of information use the publisher's licensed print and/or on-line versions! 3
potential in the treatment of Hansen's disease, atherosclerosis, immune suppression and most of all antitumor
effect [18÷50] and etc.
1.2. Clinical trial of amygdalin in the treatment of human cancer
1.2.1. Conducting research
In summary: A group of authors [33] published a detailed clinical trial of amygdalin (and its Laetrile
derivative). For the purposes of our research, the exact methodology of clinical trial must be studied in great
detail.
After fully informing the patients about the type and manner of the study, the test begins (Table 1),
constantly monitoring the concentration of total cyanide in the blood. The first analysis was performed 2 hours
after the start of oral drug administration. If the cyanide level is higher than 2 µg/ml but less than 3 µg/ml,
amygdalin is discontinued for up to 48 hours or until all symptoms suggesting toxicity are discontinued.
Therapy was continued in all patients at least until they had irrefutable evidence of progressive
malignancy or until severe clinical deterioration allowed further treatment and follow-up.
Tabl. 1. Amygdalin and “Metabolic Therapy” Regimens
AGENT
STANDARD DOSE
HIGH DOSE
Amygdalin
intravenous course
4.5 g/m2 of body-surface
7 g/m2/day X 21 days
Oral maintenance
area/day x 21 days 0.5 g 3
times daily
0.5 g 4 times daily
Vitamins
A
25,000 U/day
100,000 U/day
C
2 g/day
10 g/day
E
400 U/day
1200 U/day
B complex and minerals
1 capsule/day
1 capsule/day
Pancreatic enzymes
(Viokase)
12 tablets/day
12 tablets/day
© Charles G. Moertel, M.D., Thomas R. Fleming, Ph.D., Joseph Rubin, M.D., Larry K. Kvols, M.D., Gregory Sarna, M.D., Robert
Koch, M.D., Violante E. Currie, M.D., Charles W. Young, M.D., Stephen E. Jones, M.D., And J. Paul Davignon, Ph.D.
In short, the author's research team points out that the methods of this study are completely comparable
to those used in the studies of every new agent being developed and tested for cancer treatment through more
traditional channels. They are designed to maximize the ability of amygdalin to exhibit therapeutic activity, if
such potential exists.
1.2.2. Clinical results
The characteristics of all patients undergoing experimental treatment are listed in Table 2. Types of
tumors include predominance of colorectal, lung, and breast cancer with a standard dose regimen.
HTML 5 version of DOI: 10.2174/1871520620666200313163801 | https://anti-cancer.eu
Small typographical inaccuracies are possible. For scientific application of information use the publisher's licensed print and/or on-line versions! 4
Tabl. 2. Characteristics of eligible patients
Characteristic
High-Dose
Regimen
No. of patients
All Patients
Sex
Male
8
100
Female
6
78
Age (yr)
Median
60
57
Range
39-73
18-84
Primary tumor
Colorectal *
14
58
Lung §
30
Breast *
21
Melanoma
15
Sarcoma ,
_
10
Pancreas *
8
Stomach *
7
Kidney *
6
Lymphoma
5
Ovary *
4
Other (1 or 2 each)
14
Prior radiation therapy ֍
Yes
0
72
No
14
106(60)
Prior chemotherapy ֍
Yes
9
118
No
5
60(34)
Performance status ֍
0-1
11
127 (71)
2-3
3
51
Institution
University of Arizona
19
UCLA
40
Mayo Clinic
14
96
New York Memorial
23
* Adenocarcinoma
§ Non-small-cell carcinoma
֍ Figures in parentheses denote percentages.
Eastern Cooperative Oncology Group score: 0, fully active, to 4, totally
disabled.
© Charles G. Moertel, M.D., Thomas R. Fleming, Ph.D., Joseph Rubin, M.D., Larry K. Kvols, M.D., Gregory
Sarna, M.D., Robert Koch, M.D., Violante E. Currie, M.D., Charles W. Young, M.D., Stephen E. Jones, M.D.,
And J. Paul Davignon, Ph.D.
All patients selected for the high-dose regimen had colorectal cancer. Particularly remarkable is that
over one third of all patients did not receive prior chemotherapy. In addition, 71% of the patients had a good
working condition - that is, they were able to work full-time or part-time.
HTML 5 version of DOI: 10.2174/1871520620666200313163801 | https://anti-cancer.eu
Small typographical inaccuracies are possible. For scientific application of information use the publisher's licensed print and/or on-line versions! 5
1.2.3. Toxic reactions and symptoms
Data on the possible toxicity of intravenous and oral amygdalin are shown in Table . 3.
Tabl. 3. Toxicity of amygdalin therapy
Toxic REACTION
ROUTE
INTRAVENOUS
ORAL
% of 178 patients
% of 132 patients
Nausea
31
30
Vomiting
25
17
Headache
7
8
Dizziness
7
10
Mental obtundation
4
5
Dermatitis
2
2
© Charles G. Moertel, M.D., Thomas R. Fleming, Ph.D., Joseph Rubin, M.D., Larry K.
Kvols, M.D., Gregory Sarna, M.D., Robert Koch, M.D., Violante E. Currie, M.D., Charles
W. Young, M.D., Stephen E. Jones, M.D., And J. Paul Davignon, Ph.D.
The authors point out that adverse reactions are rare and also inherent in the oncologic diseases involved.
The side effects of increased cyanide concentration (greater than 3 μg/ml) in the blood disappear after the
amygdalin discontinuation.
Higher concentrations of amygdalin intake are also investigated, but then adverse reactions increase
significantly.
This is precisely the purpose of this article: To provide a dosage form that provides the necessary
concentration of active molecules in vivo to patients in need of anti-tumor therapy.
2. Methodology
This study is based on a hypothesis that undergoes theoretical and logical analysis to confirm and/or
reject it. The methodological scheme is:
1) Determining the optimal chemical formula - after definition, the hypothesis is subjected to two logical
proofs depending on the environment:
- in vivo - this evidence is based on a unique chemical property of the nitrile group to catalytically
convert to an amide in acidity an HCl acid. The conditions are close to those of the digestive juices
in the stomach, and the catalyst is an ion that is available under certain conditions;
- in vitro - an enzymatic reaction is analyzed to achieve the same result, i.e., modification of the
amygdalin molecule to form overlapping with the input hypothesis.
It comes with a conclusion from the part.
2) Determination of the pharmaceutical molecular form:
After confirming the defined hypothesis, it is necessary to compare the physical and chemical
relationships of the modified molecule with those of the parent amygdalin. With the help of mathematical
HTML 5 version of DOI: 10.2174/1871520620666200313163801 | https://anti-cancer.eu
Small typographical inaccuracies are possible. For scientific application of information use the publisher's licensed print and/or on-line versions! 6
chemical methods, a number of comparative methodologies are carried out, which aim to compare each of the
studied parameters between the obtained molecular forms. For this purpose, the indicators are analyzed:
- molecular topology - Molecular Topological Index, Shape Coefficient, Topological Diameter, Total
Connectivity;
- molecular networks - distribution coefficients: LogP, LogS, PKa, etc .;
- electronic reference in the atomic molecular system - Number of HBond Donors, Number of HBond
Acceptors, Formal Charge, etc .;
- molecular properties obtained by semi-empirical methods, such as: dipole moment, thermal capacity,
thermodynamic energy, etc.
Immediately after determining the average deviation between the parent and modified molecules, a
check is carried out by comparing with a well-studied, close to experimental, modification obtained by the
same methods and conditions.
It comes with a conclusion from the part.
3) Determination of the drug dose:
In order to determine the dosage, it is necessary to consider the most likely active forms of amide,
already introduced into the in vivo medium. Knowing the conditions (pH, temperature, etc.) in the human
body and comparing them with concentrations of close in molecular structure and the eventual chemical
treatment of medicinal products that have already passed clinical trials. Stoichiometric calculations are applied
and a relatively accurate dosage is obtained for the administration of the new pharmaceutical formulation in
order to maximize their anti-tumor activity.
3. Determination of the optimal chemical formula
3.1.Hypothesis
The bioactive form of amygdalin is it’s hydrolyzed to amide nitrile group.
3.2. Evidence
3.2.1. PROOF 1
Based on the chemical deposition of nitriles fig.2.
HTML 5 version of DOI: 10.2174/1871520620666200313163801 | https://anti-cancer.eu
Small typographical inaccuracies are possible. For scientific application of information use the publisher's licensed print and/or on-line versions! 7
DOI: 10.6084/m9.figshare.23898099.v1
Fig. 2. Catalytic chemical hydrolysis of nitrile to amide in an acidic environment [51]
The catalyst for this reaction can only be non-coordinatively bonded Co. Cobalt in the human body is in
the order of 22÷59 nmol/l (in the age group 30-56 years for men and women) [52]. Its daily requirement is
0.08 mg/day (for 1÷2-year-old) and reaches 0.30 mg/day (for over 60-year-old). Let us clarify that cobalamin
(known as vitamin B12) is made up of a corin ring that associates cobalt in its complexing form - therefore
cobalt in cobalamin may NOT be a catalyst for the reaction.
Based on the fact that the total content of cobalt in urine is from 3.9 to 30 nmol/d, and that of cobalamin
is 23.3 to 44.5 pmol/d, therefore there is a constant exchange of non-coordinated cobalt in the body.
After analyzing the foods consumed by Hunza people, those containing cobalt were clearly identified,
but there were no traces of vitamin B12: spinach (0.07÷1.20 mg/kg); beet leaves (0.39÷0.41 mg/kg); onions
(0.13 mg/kg); carrots (0.02 mg/kg) [53], etc.
Entered into the stomach uncoordinated cobalt (regardless of its form) will quickly dissolve under the
action of hydrochloric acid and thus enter the blood, therefore, the times of admission of amygdalin and cobalt
will not coincide. The amygdalin will enter the blood with its nitrile group from which it will release cyanide
ion.
The spinach also contains significant amounts of oxalic acid (320 mg/kg). Oxalic acid forms insoluble
cobalt oxalate (3.15 × 10
9
gr/100ml at 37 degrees of Celsius) with cobalt ions and therefore will not pass
through the gastric walls to the blood.
After analysis and simulations (based on average enthalpy and potential energy of the molecule) by TD-
DFT [54], an extremely stable retention cycle of the non-coordinated cobalt in the stomach is shown in
GAMESS US [55], shown in fig. 3:
HTML 5 version of DOI: 10.2174/1871520620666200313163801 | https://anti-cancer.eu
Small typographical inaccuracies are possible. For scientific application of information use the publisher's licensed print and/or on-line versions! 8
DOI: 10.6084/m9.figshare.23896716.v1
Fig. 3. Schematic depicting the physiological retention of cobalt in the stomach
However, at pH<2, the oxalic salts are partially hydrolyzed and metal anionic (in this case cobalt)
separated. The reaction is reversible and displaced in the direction of the starting material (cobalt oxalate).
Thus, the residence time of cobalt in the stomach increases significantly. In addition to spinach, other
cultivated plants typical of the area contain oxalic acid: tea (370 mg/kg), sorrel (360 mg/kg), rhubarb (240
mg/kg), fig (100 mg/kg), beet (40 mg/kg), plums (12 mg/kg), grapes (7 mg/kg), etc.
Under these conditions, the constantly consuming amygdalin has the conditions (concentration of
reagents, catalyst, pH, temperature, etc.) to react to catalytic hydrolysis to its corresponding amide.
3.2.2. PROOF 2
In addition to cobalt ions, the catalyst may also be cobalt linked to a covalent bond. For example, nitrile
hydratases (NHases; EC 4.2.1.84). They are metal-containing enzymes that convert the nitrile and / or cyano
group to amides, including the amygdalin fig.4.:
The optimum action of nitrile hydratase, for example from Rhodococcus rhodochrous, is from 5.5 to 8.5
pH. Their activity is highest in the range of 10÷80 degrees, and their effect is from 20 minutes to 8 hours. The
acidity in the stomach is much higher and from there these enzymes will be inhibited and even agglutinated.
HTML 5 version of DOI: 10.2174/1871520620666200313163801 | https://anti-cancer.eu
Small typographical inaccuracies are possible. For scientific application of information use the publisher's licensed print and/or on-line versions! 9
DOI: 10.6084/m9.figshare.23898177.v1
Fig. 4. Schematic diagram of the enzymatic hydrolysis of the amygdalin-nitrile group to its amides and
carboxylic acid
The peoples studied have consumed large quantities of apricot and other fruit oils. Much of the
prokaryotes and eukaryotes that produce nitrile hydratases live in lakes or in the moist environment around
them. There is a possibility, due to technological processes, to enter and live for a long time in these oils. Thus,
these insecurities may have lived in unprotected vessels and catalytically hydrolyze the amygdalin nitrile
group to amide and / or carboxylic acid.
3.3. Conclusion of the part
Regardless of the manner of preparation of the hydrolyzed nitrile group of amygdalin to amide and / or
carboxylic acid, its already modified form has a much safer chemical structure to the body. The absence of
the nitrile group, which readily converts to the cyano ion, makes it possible to increase its constant
concentration many times in the in vivo environment.
4. Determination of pharmaceutical form
All further calculations were made after optimization with respect to the minimum energy of the
molecules via MM2 [56] and MMFF94 [57] and followed by molecular dynamics responsible for 310 K, 1.02
bar in a medium of 0.9% solution of NaCl in water using CPCM [58] methodology.
HTML 5 version of DOI: 10.2174/1871520620666200313163801 | https://anti-cancer.eu
Small typographical inaccuracies are possible. For scientific application of information use the publisher's licensed print and/or on-line versions! 10
4.1. Mathematical Chemical Indicators
4.1.1. Molecular topology
For a more complete characterization of the hypothetical molecules, they were compared with the
starting one (amygdalin). Comparisons were made with respect to: molecular topology (Table 4), Molecular
Networks (Table 5), electronic reference in the atomic-molecular system (Table 6), and physical and
thermodynamic parameters (Table 7).
Tabl. 4. Comparative analysis of the molecular topology of pure amygdalin, its amide and carboxyl
derivatives, resulting from the hydrolysis of its nitrile group
-CN
-C(O)NH
2
-COOH
Balaban Index [59÷63]
1660564
1893740
1893740
Cluster Count
32
33
33
Molecular Topological Index
20391
21571
21342
Num Rotatable Bonds, [No. of bonds]
8
8
8
Polar Surface Area [Å
2
]
202.32
221.62
215.83
Radius, atoms
8
8
8
Shape Attribute
30.03125
31.03(03)
31.03(03)
Shape Coefficient
0
0
0
Sum Of Degrees
68
70
70
Sum Of Valence Degrees
124
128
130
Topological Diameter
15
15
15
Total Connectivity
2.14E-05
1.75E-05
1.75E-05
Total Valence Connectivity [64]
8.47E-10
4.46E-10
3.46E-10
Wiener Index [65,66]
3080
3308
3308
4.1.2. Molecular Networks:
Tabl. 5 Partition coefficients of amygdalin and its hydrolysates of the nitrile group to amide and carboxylic
acid in the 0.9% NaCl phase in water
-CN
-C(O)NH
2
-COOH
LogP [67]
-2.81
-3.67
-2.93
LogS [68]
-0.047
0.216
-0.036
PKa [69]
O-1
16.644
16.7357
18.794
O-2
17.4061
17.4061
17.5938
O-3
12.889
12.9592
13.0704
O-4
13.0331
13.033
13.0254
O-5
18.9903
19.0068
16.983
O-6
20.6647
20.6646
20.6647
O-7
14.9367
14.9366
14.9367
HTML 5 version of DOI: 10.2174/1871520620666200313163801 | https://anti-cancer.eu
Small typographical inaccuracies are possible. For scientific application of information use the publisher's licensed print and/or on-line versions! 11
OH(COOH)
3.03401
Molar Refractivity [70]
10.525
10.915
10.701
4.1.3. Electronic treatment in atomic molecular system
Tabl. 6. Electron Assignment in the Atomolecular System [71,72] of amygdalin and its hydrolysates of the
nitrile group to amide and carboxylic acid
-CN
-C(O)NH
2
-COOH
Formal Charge
0
0
0
Number of HBond Acceptors
12
12
12
Number of HBond Donors
7
8
8
Ovality
NaN
NaN
NaN
Principal Moment
1901.916
6152.891
6968.502
2042.063
6108.058
7033.240
2100.609
6066.442
7045.560
Henry's Law Constant [73]
24.508
27.945
26.255
4.2. Molecular properties obtained by semi-empirical methods
Tabl. 7. Molecular and electron-configuration properties of amygdalin and its hydrolysates of the nitrile
group to amide and carboxylic acid obtained by semi-empirical methods [74]
method
-CN
-C(O)NH
2
-COOH
Core-Core Repulsion, [eV]
PM7 [75]
51868.25
56375.19
56412.48
COSMO Area, [Å
2
]
378.83
385.94
387.52
COSMO Volume, [Å
3
]
489.25
508.77
507.20
Dipole, [Debye]
4.991
5.84
5.71
Ionization Potential, [eV]
9.93
9.87
9.90
Total Energy, [eV]
-6254.95
-6578.48
-6674.00
Heat Capacity, [Cal/Mol-Kelvin]
PM6 [76]
/ 310.15 K,
1.02 bar /
96.27
100.19
97.03
Thermodynamic Energy, [Kcal/Mol]
283.95
297.77
289.42
Data in Tables 4, 5, 6, 7 show that values (including their equated statistical error) over 23% of the
indicators are the same. No drastic topological and molecular network deviations and electron-configuration
anomalies are observed. The differences represent up to 12% mean deviation of the amygdalin index with its
hydrolyzed derivatives.
HTML 5 version of DOI: 10.2174/1871520620666200313163801 | https://anti-cancer.eu
Small typographical inaccuracies are possible. For scientific application of information use the publisher's licensed print and/or on-line versions! 12
To test the tolerance of 12%-th still not jeopardize the normal physiology of the body, let us consider a
well-studied enzyme hydrolytic industrial process (fig.5) of a passage of 3-Cyanopyridine to Nicotinamide
[77] (vitamin B3) by means of nitrile hydratase. The thermodynamic quantities are compared: total energy,
thermal capacity, and thermodynamic energy, since these physical functions are the consequence of most
topological, morphological, and electron-configuration relationships of each molecule in tabl.5.
DOI: 10.6084/m9.figshare.23898204.v1
Fig. 5. Enzyme hydrolysis of 3-Cyanopyridine to Nicotinamide and nicotinic acid
It is important to note that all three enzymes are almost always together, whether of micro-bacterial or
bio-physicochemical origin.
Tabl. 5. Total Energy, Heat Capacity and Thermodynamic Energy on 3-Cyanopyridine, Nicotinamide and
Nicotinic Acid by 310 K, 1.02 bar in medium of 0.9% NaCl solution in water using CPCM [78] methodology
3-Cyanopyridine
Nicotinamide
Nicotinic
acid
Total Energy PM7, [eV]
-1162.27
-1485.64
-1581.25
Heat Capacity, [Cal/Mol-Kelvin]
16.73
19.75
20.37
Thermodynamic Energy, [Kcal/Mol]
55.72
70.77
63.54
By comparing the data in Table 5 with the conclusions of Table 1, 2, 3, 4, the average standard deviation
is confirmed when comparing the molecular nature of the nitriles, the amide and their corresponding
carboxylic acid.
4.3. Conclusion of the part
Due to the theoretical molecular modification, namely the transition from condensed and / or
polymerized nitrile to carbohydrate in its amide and as a by-product and its acid, does not change the total
activity of the parent compound as well as its active groups not involved in hydrolysis. The chemical stability
and predictability of hydrolysis products are the basis for a pharmaceutical formulation that covers all
international requirements for a conservative medicinal product.
HTML 5 version of DOI: 10.2174/1871520620666200313163801 | https://anti-cancer.eu
Small typographical inaccuracies are possible. For scientific application of information use the publisher's licensed print and/or on-line versions! 13
5. Determine drug dose
In order to determine the dosage, it is necessary to consider the most likely active forms of amide already
in vivo.
In oral use (which is also recommended), the connection between the two glycosidic nuclei will be
attacked by the salivary gland. Due to the relatively short time for complete contact between the substance
and the enzyme, the probability of a reaction being below 3% and