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Theoretical study of the process of passage of glycoside amides through the cell
membrane of cancer cell
Vasil Tsanov, Hristo Tsanov
ABSTRACT
Background: This article concentrates on the processes occurring in the medium around the cancer cell and
the transfer of glycoside amides through their cell membrane. They are obtained by modification of natural
glycoside-nitriles (cyano-glycosides). Hydrolysis of starting materials in the blood medium and associated
volume around physiologically active healthy and cancer cells, based on quantum-chemical semi-empirical
methods, is considered.
Objective: Based on the fact that the cancer cell feeds primarily on carbohydrates, it is likely that organisms
have adapted to take food containing nitrile glycosides and / or modified forms to counteract "external"
bioactive activity. Cancers, for their part, have evolved to create conditions around their cells that eliminate
their active apoptotic forms. This is far more appropriate for them than changing their entire enzyme regulation
to counteract it. In this way, it protects itself and the gene sets and develops according to its instructions.
Methods: Derived pedestal that closely defines the processes of hydrolysis in the blood, the transfer of a
specific molecular hydrolytic form to the cancer cell membrane and with the help of time-dependent density-
functional quantum-chemical methods, its passage and the processes of re-hydrolysis within the cell itself, to
forms causing chemical apoptosis of the cell - independent of its non-genetic set, which seeks to counteract
the process.
Results: Used in oncology it could turn a cancer from a lethal to a chronic disease (such as diabetes). The
causative agent and conditions for the development of the disease are not eliminated, but the amount of cancer
cells could be kept low for a long time (even a lifetime).
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Conclusion: The amide derivatives of nitrile glycosides exhibit anti-cancer activity; the cancer cell probably
seeks to displace hydrolysis of these derivatives in a direction that would not pass through its cell membrane
and the amide-carboxyl derivatives of nitrile glycosides could deliver extremely toxic compounds within the
cancer cell itself and thus block and / or permanently damage its normal physiology.
Keywords
glycoside amides, cancer cell membrane, hydrolysis, PM7, TD-DFT, apoptosis
1. Background
This scientific analysis is a continuation of the article Theoretical analysis for the safe form and dosage
of amygdalin product [1]. The hypothesis that hydrolyzed to amine / carboxylic acid cyano / nitrile glycosides
are a potential anti-cancer drug has been proposed and theoretically confirmed there. Their biological activity
remains unchanged directly from the natural compounds of this group, but their toxicity is many times lower
than unmodified native molecules. After defining the chemical formula and determining the pharmaceutical
form and dosage, most active groups are also identified, which directly determines their biological activity,
schematically represented in Fig. 1.
DOI: 10.6084/m9.figshare.23898555.v1
Fig. 1. Summary scheme of the theoretically calculated anticancer activity of the biologically modified
amygdalin and the site of the dosage form throughout the biochemical cycle
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The proposed pharmaceutical form of the amide / carboxyl derivative of amygdalin represents only one
of the dozens of glycoside nitriles that have been analyzed and by which this claim is made. Some of them are
listed [2, 3] in Table 1.
Tabl. 1. Natural nitrile glycosides and their modified forms
Base Structure
Substituent R
Glycoside
Sugar
Occurrence
Glycosides with aromatic substituents
Phenyl
Prunasin
D-Glucose
Prunus spp.
Phenyl
Amygdalin
Gentiobiose
Prunus spp.
Phenyl
Lucumin
Primeverose
Lucuma spp.
Phenyl
Vicianin
Vicianose
Vicia spp.
Phenyl
Sambunigrin
D-Glucose
Sambucus spp.
p - Hydroxyphenyl
Dhurrin
D-Glucose
Sorghum spp.
p - Hydroxyphenyl
Taxiphyllin
D-Glucose
Taxus spp.
m - Hydroxyphenyl
Zierin
D-Glucose
Zieria spp.
p - Glucosyloxyphenyl
Proteacin
D-Glucose
Macadamia spp.
Glycosides with a free α - hydroxynitrile
p-Glucosyloxymandelonitrile
Nandina spp.
Glycosides with Aliphatic substituents
R=R’=CH
3
Linamarin
D-Glucose
Linum spp.
Trifolium spp.
R=CH
3
R’=CH
2
.CH
3
Lotaustralin
D-Glucose
Loyus spp.
Maniholt spp.
R=C(CH
3
)
2
Acacipetalin
D-Glucose
Acacia spp.
R=C(CH
2
.COOH).CH=
CH-COOH
Triglochinin
D-Glucose
Triglochin spp.
R=R
’
=H
Deidaclin
D-Glucose
Deidamia spp.
Tetraphyllin A
D-Glucose
Tetrapathaea
spp.
R=OH; R’=H
Tetraphyllin B
D-Glucose
Tetrapathaea
spp.
R=R’=OH
Gynocardin
D-Glucose
Gynocardia spp.
Pangium spp.
It is important to mention that the by-product of the modification process also produces their carboxyl
derivatives in a ratio, the ratio for amygdalin being: -amide: -carboxyl = 4.87: 1, and for other homologs and
/ or similar structural of compounds is in the order of 2.61 ÷ 5.13: 1. Chemical bond of the type: -N(H)-OC(O)
- between the two derivatives of amygdalin is possible, but it is within statistical error.
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2. Objective
Under standard physiological conditions in the body, the presence of a carboxyl derivative is not a
hindrance factor, but on the contrary, it would stabilize the reactions of the amido product of amygdalin to
some extent, due to its well-expressed proton activity.
The present study is based on the scientific fact that a relatively conditioned associated volume around
the cancer cell [4] has a low acidity [5] (pH = 6.5, at a physiologically healthy cell norm pH = 7.4). This
circumstance, from the point of view of the precision of quantum-chemical and molecular-topological [6]
methods, makes it possible to carry out a sufficiently reliable comparative analysis [7] of substances in the
immediate vicinity of the cells and to differentiate their chemical relations in the respective media.
Semi-empirical methods [8, 9] are extremely suitable when comparing individual parameters directly
related to the chemical and bioactive properties of individual molecules. They do not claim accuracy, but give
a fairly clear picture when comparing individual calculated and / or measured values, and are sufficiently
reliable, especially for identifying them in individual structural relationships.
3. Methodology
Conditions selected to be maximal in vivo: are at temperature 37 degrees Celsius, in blood medium
(reported by the dielectric constant, not entering this indicator into the equations results in a deviation of 5 to
8% - which is not physiologically justified).
The dialectic constant of human blood has a different value in different blood groups [10, 11], so we take
its average arithmetic as a starting point – 68.15 x10
3
at 37
o
C and at the frequency of 1 KHz.
The electrostatic potential, created in the space around a molecule [12] of its nuclei and electrons (treated
as static charge distributions), is a well-applied property for analyzing and predicting molecularly reactive
behavior. Hence the approximation that the molecular electrostatic potential is the potential energy of a proton
at a specific location near a molecule. In this case, it is particularly useful as an indicator of the sites or zones
of a molecule, initially attracted by an approaching electrophile, and has also been successfully applied to
investigate interactions, that involve a certain optimal relative orientation of the reagents, in our case as an
active pharmaceutical form cell to pass a specific cell membrane (cancer). This, however, prevents the
recording of charges originating from the local electron density (charges density), which could be achieved
with Mulliken Changes. The use of both variables from different methodologies more objectively illustrates
the charges in the molecule and facilitate subsequent interpretation. When comparing the outputs, it is
necessary to use the same functional and basic set to make an accurate comparison, since the electron density
is sensitive precisely to the likelihood of the process (whether at the electrostatic level).
With the introduction of ionization potential, as a corrective for the interpretation of individual factors
in the construction of the overall picture, more dualistic interpretation values (including variables) are
eliminated.
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All other investigated indicators are introduced into the analysis in order to better characterize the
processes and their approximation to the actual patho & sanus physiological environment in the body.
The whole presentation is based on the amide and carboxyl form of amygdalin as the best studied, but
they are only one representative of the homologous order. The conclusions are absolutely comparable to all
its representatives (in their respective relation - admin to carboxylic acid §1.).
Each source molecule is pre-exposed to MM2 [13] minimized energy, followed by molecular dynamics
simulation at 37
o
C (with 100 iterations) in order to generate an input to the subsequent Z-matrix [14]
calculations. All values obtained by semi-empirical methods were performed in MOPAC [15] environment
with type PM7 [16] and those of time-dependent density-functional theory (TD-DFT) in a GAMESS US [17]
environment. Each value is calculated five times with five different computer configurations and OS (Linux -
Ubuntu™ & OpenSuse™; Windows™ - 7 Pro & 10 Workstation). The results were statistically analyzed by
the mean squared error [18] and the hypotheses with the Student's T-test [19] for independent samples. The
final numeric value is the one closest to its neighbor (also represented).
3.1.Characterization of amide dissociation in vivo
Analyzes the non-hydrolyzed amide / Basic Form A - BF(A) / both hydrolytic forms / conditionally
accepted on Hydrolytic Form A - HF(A) and Hydrolytic Form B - HF(B) / in steps according to the indicators:
▪ Electrostatic Potential [20] (of reactive atoms), Mulliken Changes [21], Core-Core Repulsion [22],
COSMO [23, 24] Area [25] and Volume [26], Dipoles [27, 28] (vector Debye), Electronic & Total Energy
and Ionization Potential [29];
▪ Polar Surface Area [30], Radius [31] and Topological Diameter [32];
▪ LogP, LogS and Partition Coefficient [33, 34];
▪ calculation and structural representation of pKa [35, 36] of each atom and / or group of the whole
molecule.
To carry out the check is applied and Principal Moment [37] and Lipinski Rule [38] (etc. Lipinski's rule
of five).
3.2. Characterization of the carboxylic acid obtained as a by-product of nitrile hydrolysis
It shall be performed identically according to §3.1.
3.3. Schematic presentation of the data from §3.1. and §3.2. with the necessary interpretations and
conclusions - with the title: Mechanism of penetration of the modified molecule into the cancer cell.
TD-DFT is applied with respect to potential energy [39] and average enthalpy [40].
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4. Results and analysis
4.1. Characterization of amide dissociation in vivo
For the reaction-determining atoms, the following were calculated: Mulliken Changes, Electrostatical
Potential, Core-Core Repulsion, COSMO Area and Volume, Dipoles (vector Debye), Electronic & Total
Energy and Ionization Potential.
Mulliken Changes and Electrostatical Potential of the reactive atoms in amide derivative of amygdalin
and its two hydrolytic forms in vivo are shown in Fig.2.
DOI: 10.6084/m9.figshare.23899164.v1
Basic form A
Hydrolytic Form A
Hydrolytic Form B
black – Mulliken Changes
red – Electrostatical Potential
Fig. 2. Mulliken Changes and Electrostatical Potential of the amide derivative of amygdalin and its two
hydrolytic forms in vivo
The negative electrostatic potential corresponds to the attraction of the proton from the electron density
concentrated in the geometric space in the molecules (from single pairs, π-bonds, etc.). The positive
electrostatic potential corresponds to the repulsion of the proton from the atomic nuclei in regions with low
electron density and the nuclear charge is incompletely shielded. The data of Fig.2 clearly illustrates that
during hydrolysis of glycoside amide, obtained by hydrolytic modification of nitrile glycoside (in this case
amygdalin) in vivo, the electrostatic equilibrium shifts to the ammonia-saturated form (assumed conditionally
for Hydrolytic Form A). This is confirmed by Mulliken Changes [41] of the three molecular forms.
Therefore, in terms of Electrostatical Potential and Mulliken Changes, the medium around the
cancer cell tends to shift the hydrolytic equilibrium to HF(A). This is provided that the molecule is relatively
static and already reoriented in a closed volume around the cancer cell.
The Core-Core Repulsion, COSMO Area and Volume, Electronic Energy, Ionization Potential and Total
Energy of the reactive atoms in amide derivative of amygdalin and its two hydrolytic forms in vivo are depicted
in Fig.3.
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DOI: 10.6084/m9.figshare.23899782.v1
Fig. 3. Core-Core Repulsion, COSMO Area and Volume, Electronic Energy, Ionization Potential and Total
Energy of amygdalin amide derivative and its two hydrolytic forms in vivo
Core-Core Repulsion has the highest value at HF(A). This is an indicator of a slight reorientation of the
proton centers, which is confirmed by the slight geometric deformation observed in this form. COSMO Area
and Volume grow in the direction from the ammonia-saturated to the hydroxy-saturated hydrolytic molecular
form. These circumstances indicate that HF (B) occupies a larger geometric volume (along with associated
blood / water / intercellular fluid / CSF), which minimizes the shielding of externally charged molecules
(including dipoles, ions, domains, etc.). This is confirmed by the lower ionization energy compared to HF(A).
This effect is not decisive in the expression of total energy (i.e. total energy affects ionization potential, not
the other way around). It is apparently lower than BF(A), and is relatively the same in HF(A) and HF(B).
Therefore, in terms of Core-Core Repulsion, COSMO Area and Volume, Electronic Energy,
Ionization Potential and Total Energy, the environment around the cancer cell tends to support hydrolysis
to HF(B).
There is a certain dualism in this summary: Provided that the reorientation in space is due to the
elimination of energy and / or partial charges, and they in turn increase the polar hydration volume (an integral
part of the total volume of the molecule), non-uniform polarization inside the basic structure of the molecule
under relatively constant conditions. The latter is an indisputable fact, since the in vivo medium is buffered to
both pH and temperature.
The polarizations within the molecular forms can be compared by so-called Debye vectors on the
molecular spatial axes along the X, Y, Z and total dipole moment of the amide derivative of amygdalin and
its two hydrolytic forms in vivo are shown in Fig. 4.
DOI: 10.6084/m9.figshare.23900082.v1
Basic Form А Hydrolytic Form A Hydrolytic Form B
370
372
374
376
378
380
382
384
386
388
390
392
394
396
398
400
402
404
COSMO Area
COSMO Volume
Core-Core Repulsion
Electronic Energy
Ionization Potential
Total Energy
480
490
500
510
520
530
540
6760
6780
6800
6820
6840
6860
6880
6900
6920
6940
6960
6980
7000
7020
7040
-13840
-13820
-13800
-13780
-13760
-13740
-13720
-13700
-13680
-13660
-13640
-13620
-13600
-13580
-13560
-13540
-13520
9,9
10,0
10,1
10,2
-6592
-6590
-6588
-6586
-6584
-6582
-6580
A
3
A
2
eV
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Fig. 4. Dipole moment of amygdalin amide derivative molecule and its two hydrolytic forms in vivo
HF(A) has a lower dipole moment, with the same molecular radius, topological diameter, and polar
surface area (Table 2). The analysis thus performed does not take into account the change of the dipole moment
under the dynamic action of the active molecules.
Tabl. 2. Polar Surface Area, Molecular Radius and Topological Diameter of amygdalin amide derivative
molecule and its two hydrolytic forms in vivo
Basic
Form A
Hydrolytic
Form A
Hydrolytic
Form B
Molecular Topology
Polar Surface Area
Å
221±4
223±4
226±4
Radius
Atom(s)
8
8
8
Topological Diameter
Bond(s)
15
15
15
This circumstance does not allow us to assume the common belief that the lower dipole moment has
lower molecular activity in a polar environment. Therefore, we need to analyze the partition coefficients pKa
(Fig.5), LogP, LogS and Partition Coefficient (Table 3) in the starting conditions and conditions (§3.).
The calculation and structural representation of pKa of each atom and / or group of the entire molecule
in all three forms is presented in Fig.5. Black values are unchanged during hydrolysis, red those that increase
and blue are those that decrease.
DOI: 10.6084/m9.figshare.23900118.v1
Basic Form A Hydrolytic Form A Hydrolytic Form B
-5
0
5
10
15
X
Y
Z
Total
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Fig. 5. pKa per atom and / or group of the whole molecule in amide of both its hydrolytic forms of
amygdalin
Based on the definition of pKa, namely that it is inversely proportional to the "strength" of the acid (or
acid residue) and the degree of more complete dissociation, it follows that HF(A) is more alkaline than HF(B),
relative to the major hydrolytically active groups (-[C=O]-NH
3
+
и –[C=O
+
H]-NH
2
).
HF(B) has a change in the hydrolytically active hydroxyl groups of the glucosidal nuclei. From thence,
the internal polarization of the molecule also grows, which is also confirmed by the higher dipole moment
(Fig.4) with respect to HF(A). It is important to note that this does not compare the dipole moments in the
molecule itself with that of the molecule and the solvated volume around it. These are two different dimensions
in which external factors are crucial. In our case, even with constant temperature and acidity, the molecule is
also affected by other ions and molecules, and in some cases by mechanical effects (including movement of
blood, cerebrospinal fluid, lymph, tissue and intercellular fluids).
In terms of distribution coefficients, both hydration forms are equally likely to occur. On the one hand,
the more acidic medium will shift the equilibrium to ammonia-saturated HF(A), and on the other hand, the
speed and stability of hydrolysis will be faster on HF(B) - probably due to incomplete protonation by the more
acidic medium around the cancer cell. The data from Principal Moment (Table 3) cannot be interpreted
uniquely.
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Tabl. 3. Principal Moment, Lipinski Rule of Five, LogP, LogS, Partition Coefficient of amygdalin amide
derivative
Basic
Form A
Hydrolytic
Form A
Hydrolytic
Form B
Principal Moment
2743
4688
5994
2711
4606
5876
2824
4484
5910
Lipinski Rule of Five
molecular weight
475.169
476.176
476.176
number of HBond
acceptors
units
12
12
11
number of HBond
donors
units
8
8
9
number of rotatable
bonds
units
8
8
8
LogP
Log Units
-2.976
-2.673
-2.673
LogS
Log Units
0.216
0.204
0.934
Partition
Coefficient
conditional
units
-2.976
-2.673
-2.673
There is a deviation from the Lipinski Rule of Five application indicator. This is due to the molecular
topology of the three forms - BF(A) and HF(A; B). The molecules are divided into four major parts - two
Substituent (or H), Sugar (Fig.1) and an amide group. They have different electron density (and / or protons)
transported in different directions, with different capacities and intensities. The very algorithm for deriving
the Lipinski Rule of Five is quite subjective and with a few variables that we eliminate through analysis.
Behind each statement is at least three counter-analyzes and the necessary statistical processing (according to
the methodology – §3.).
Conclusion of the part:
Based on the aforementioned theoretical-analytical considerations, it can be concluded that the cancer
cell seeks to maintain hydrolysis to HF(B). Following the same analysis using a completely identical
methodology, but at pH=7.4, it is concluded that a physiologically sound cell tends to undergo more alkaline
hydrolysis, namely HF(A).
As mentioned in §1., in addition to the amide derivative, there are invariably significant amounts of the
corresponding carboxylic acid / BF(C) / with it. This compound (even at low concentrations) plays a
significant role in the ionic activity of the medium and hence the chemical activity of the hydrolyzed amide /
BF(A) /.
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4.2. Characterization of carboxylic acid dissociation in vivo
For the reaction-determining atoms, the following were calculated: Mulliken Changes, Electrostatical
Potential, Core-Core Repulsion, COSMO Area and Volume, Dipoles (vector Debye), Electronic & Total
Energy and Ionization Potential.
The Mulliken Changes and Electrostatical Potential of the reactive atoms amide derivative of amygdalin
and its two hydrolytic forms in vivo are shown in Fig.6.
DOI: 10.6084/m9.figshare.23900181.v1
Basic form C
Hydrolytic Form C
black – Mulliken Changes
red – Electrostatical Potential
Fig.6. Mulliken Charges and Electrostatic Potential of the carboxyl derivative of amygdalin and its
hydrolytic forms in vivo
According to the analytical wording in §4.1. concludes that the hydrolytic equilibrium of the reaction is
shifted from BF(C) to HF(C). The difference in ionic activity at pH = 6.5 and 7.4 is within less than 1%
deviation (due to the presence of only one hydrolytic form), it is evident that the values are in the statistical
error of the methods.
Therefore, from the perspective of Electrostatical Potential and Mulliken Changes, the medium
around the cancer cell tends to shift the hydrolytic equilibrium to HF(C). The influence of the dynamic
relationships of the molecule is not essential for hydrolysis around healthy and cancer cells.
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DOI: 10.6084/m9.figshare.23900208.v1
Fig. 7. Core-Core Repulsion, COSMO Area and Volume, Electronic Energy, Ionization Potential and
Total Energy of amygdalin carboxyl derivative and its hydrolytic forms
in vivo
The data of Fig.7, Fig.8 and Table 4 shall be interpreted in proportion to what the data of §4.1.
DOI: 10.6084/m9.figshare.23903694.v1
Fig. 8. Dipole moment of the carboxyl derivative of amygdalin and its hydrolytic forms in vivo
Basic Form C Hydrolytic Form C
370
372
374
376
378
380
382
384
386
388
390
392
394
396
398
400
402
404
406
408
COSMO Area
COSMO Volume
Core-Core Repulsion
Electronic Energy
Ionization Potential
Total Energy
480
490
500
510
520
530
540
6900
6800
6700
6600
6500
6400
6300
-12900
-13000
-13100
-13200
-13300
-13400
8,0
8,5
9,0
9,5
10,0
-6700
-6650
A
3
A
2
eV
Basic Form C Hydrolytic Form C
-30
-25
-20
-15
-10
-5
0
5
10
15
20
25
30
35
X
Y
Z
Total
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Tabl. 4. Polar Surface Area, Molecular Radius and Topological Diameter of carboxyl derivative of
amygdalin and its hydrolytic forms in vivo
Basic Form C
[pH=7.4]
Hydrolytic Form C
[pH=6.5]
Molecular Topology
Polar Surface Area
Å
216±4
225±4
Radius
Atom(s)
8
8
Topological Diameter
Bond(s)
15
15
The pKa of the starting material BF(C) and the reaction product HF(C) in this case are not significant
when considering the reaction. This is confirmed by the same Polar Surface Area (Table 4) for the molecular
forms in the two acidic states. The activity of the hydroxyl groups in the non-structural position will assume
a value close to that of the natural nitrile. In order not to get an active chemical form that is completely different
in bio-reactivity, the starting carboxylic molecule is also compared with respect to Principal Moment, Lipinski
Rule of Five and the partition coefficients (Table 5). The deviation from them is comparable to that of the
amide form (§4.1.).
Tabl. 5. Principal Moment, Lipinski Rule of Five, LogP, LogS, Partition Coefficient of amygdalin
carboxyl derivative
Basic form C
Hydrolytic Form C
Principal Moment
2833
4587
5953
2881
4321
5827
Lipinski Rule of Five
molecular weight
g/mol
476.153
476.153
number of HBond acceptors
numbers
12
13
number of HBond donors
numbers
8
7
number of rotatable bonds
numbers
8
8
LogP
Log Units
-2.934
-3.109
LogS
Log Units
-0.036
0.8123
Partition Coefficient
conditional
units
-2.080
-10.548
Conclusion of the part:
Based on the analysis, we conclude that acidity around the cancer cell is not a significant factor affecting
hydrolysis. It is again moved in the direction of the product, i.e. HF(C).
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5. Mechanism of penetration of the modified molecule into the cancer cell
The biological activity of the hydrolyzed to amide / carboxylic acid nitrile / cyano glycosides (Table 1)
is approximately the same as that of the native (basic) molecules. This allows high doses (§1.) to be
administered, which in turn significantly increases their anticancer activity (in terms of concentration).
In Fig.9 is a general schematic view of the data of Table 1 and their hydrolysis [42].
DOI: 10.6084/m9.figshare.23903712.v1
Fig. 9. Hydrolysis of amide and carboxylic acid derivatives of nitrile (cyano) glycosides
Figure 10 presents two physiologically active cells: healthy (III.) with pH=7.2 and cancer (V.) with
pH=7.4. They are found in a volume of blood (I.) with pH=7.4. Around each cell has an associated volume of
liquid (VI.) With specific ionic activity and hence different acidity. For a healthy cell, pH=7.4 and for cancer
pH=6.5. Based on the data of it.3, it follows that in a healthy cell, all three hydration forms HF(A;B;C) will
be in significant concentration both in the blood (I.) and in the closed volume (II.) around it. In the volume
around the cancer cell, the hydrolytic equilibrium is shifted in the HF direction (B). In this form, the molecule
loses its activity in an environment of excess protons, i.e. behaves like a "regular carbohydrate".
Based on the fact that the cancer cell feeds primarily on carbohydrates, it is likely that the organisms
have adapted to receive food containing nitrile glycosides and / or their modified forms to counteract
"external" biological effects. Cancers, for their part, have evolved to the extent that they create conditions
around their cells that eliminate the active apoptotic forms. This is far more appropriate for them than changing
their entire enzyme regulation to counteract it. In this way, it protects itself and the gene set and develops
according to its instructions.
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DOI: 10.6084/m9.figshare.23903739.v1
Fig. 10. Schematic representation of the type of hydrolysis of amidе and carboxylic acid derivatives of
nitrile (cyano) glycosides around a physiologically active cancer and healthy cell
Therefore, the hydration balance of HF(A,B) in the blood, the medium around the cancer cell shifts it in
the direction of HF(B). Parallel to this hydrolysis, the carboxylic acid, i.e. from BF(C) to HF(C). It is not
sensitive to this change in acidity and the equilibrium is shifted towards the product. This concludes that the
concentration of HF(C) is approximately the same in (I) and (IV.) in healthy and cancer cells. This form also
could hardly pass through the cell membrane in considerable concentration.
The presence of both types of hydrolysates in one volume in the blood (I.) changes the whole picture.
The equilibrium at the amide derivative is shifted (Fig.11) in direction HF(A).
DOI: 10.6084/m9.figshare.23903775.v1
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Fig. 11. Formation of a complex with incomplete counter-charge between hydrolysis of amide and
carboxylic acid derivatives of nitrile (cyano) glycosides
The compound thus obtained exhibits hydrolytic inertness at pH=6.4. Thus, the resulting form HF(A;C)
reaches unaltered to the cell membrane of the cancer cell. These compounds are due to the sharing of
incomplete electron charges and are stabilized by a solvate shell of water. The shortage of OH groups further
stabilizes the process, due to the slightly acidic medium and free protons oriented directly above the cell
membrane. In parallel with this process, reverse hydrolysis takes place (Fig.12) from HF(C) to BF(C) - which
retains activity and leaves the associated volume around the cancer cell, re-hydrolyzes and binds a new amount
of HF(A) and enters into the closed volume.
DOI: 10.6084/m9.figshare.23903805.v1
Fig. 12. Scheme of chemical bonding between basic and hydrolyzed forms of amide and carboxyl forms of
nitrile glycosides under various conditions in vivo around and in cancer cell
The HF(A) molecule contains at least one glycosidic group. The compound is saturated with ammonia
(-NH
3
+
) and readily binds with protein to glycoprotein. The hydrolysis-modified form is passed through a
protein carrier through the cell membrane. Here, however, pH is equal to 7.4 and saturation of -OH groups.
Thus, HF(B) is obtained, i.e. there is a partial shift of the hydrolytic equilibrium from HF(A) through BF(A)
to HF(B). An enzyme amidase [43] is also synthesized in the cell, which converts -(CO).NH
2
to -(COOH). As
a final product under these conditions we have all three hydrolytic forms - HF(A;B;C).
Therefore, the eventual chemical apoptosis will proceed independently of all enzymes synthesized
according to instructions from cancer DNA (for example, the linamarase gene to linamarase).
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6. Toxication of the cancer cell
Active apoptotic forms (AAF) with manifested anti-cancer activity are formed according to their
molecular structure. For diglycoside compounds (Amygdalin / Gentiobiose /, Lucumin / Primeverose /,
Vicianin / Vicianose /, etc.) primary enzymatic hydrolysis (gluconases - which are abundant in tissue fluids)
of the glycosidic bonds between the individual sugars takes place. The relationship between the secondary
carbohydrate and the reaction-determining group is stronger and requires a longer reaction time and / or a
specific enzyme such as amygdalin beta-gluconase. The latter is synthesized mainly inside the cell itself. This
leads us to conclude that the passage through the cell membrane of the cancer cell (§5.) occurs with only one
carbohydrate molecule. Once inside the cell, the only glycosidic bond is broken. This is how the AAFs
themselves are created. Some of them are listed in Table 6.
Tabl. 6. Active apoptotic amide / carboxyl molecular forms
chemical formula
designations
natural precursor
(R)-2-hydroxy-2-phenylacetamide
Prunasin
Amygdalin
Lucumin
Vicianin
Sambunigrin
(R)-2-hydroxy-2-phenylacetic acid
(R)-2-hydroxy-2-(4-
hydroxyphenyl)acetamide
Dhurrin
Taxiphyllein
Proteacin
p-Glucosyloxymendelonitrile
(R)-2-hydroxy-2-(4-
hydroxyphenyl)acetic acid
(R)-2-hydroxy-2-(3-
hydroxyphenyl)acetamide
Zierin
(R)-2-hydroxy-2-(3-
hydroxyphenyl)acetic acid
2-hydroxy-2-methylpropanamide
Linamarin
2-hydroxy-2-methylpropanoic acid
(S)-2-hydroxy-2-methylbutanamide
Lotaustralin
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(S)-2-hydroxy-2-methylbutanoic acid
2-hydroxy-3-methylbut-2-enamide
Acacipetalin
2-hydroxy-3-methylbut-2-enoic acid
(2Z,4E)-4-(2-amino-1-hydroxy-2-
oxoethylidene)hex-2-enedioic acid
Triglochinin
(2E,4Z)-3-(carboxymethyl)-2-
hydroxyhexa-2,4-dienedioic acid
(S)-1-hydroxycyclopent-2-ene-1-
carboxamide
Deidaclin
Tetraphyllin A
(S)-1-hydroxycyclopent-2-ene-1-
carboxylic acid
(1S,4S)-1,4-dihydroxycyclopent-2-
ene-1-carboxamide
Tetraphyllin B
Volkenin
Taraktophyllin
(1S,4S)-1,4-dihydroxycyclopent-2-
ene-1-carboxylic acid
(1R,4R)-1,4,5-trihydroxycyclopent-2-
ene-1-carboxamide
Gynocardin
(1R,4R)-1,4,5-trihydroxycyclopent-2-
ene-1-carboxylic acid
(Z)-2-((4S,6R)-4,6-
dihydroxycyclohex-2-en-1-
ylidene)acetamide
Menisdaurin
(Z)-2-((4S,6R)-4,6-
dihydroxycyclohex-2-en-1-
ylidene)acetic acid
(R)-2-hydroxy-3-methylbutanamide
Volkenin
(R)-2-hydroxy-3-methylbutanoic acid
(E)-2-((4S,5R,6R)-4,5,6-
trihydroxycyclohex-2-en-1-
ylidene)acetamide
Griffonin
(E)-2-((4S,5R,6R)-4,5,6-
trihydroxycyclohex-2-en-1-
ylidene)acetic acid
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(Z)-2-((4R,5R,6S)-5,6-dihydroxy-4-
methoxycyclohex-2-en-1-
ylidene)acetamide
Bauhinin
(Z)-2-((4R,5R,6S)-5,6-dihydroxy-4-
methoxycyclohex-2-en-1-
ylidene)acetic acid
(E)-2-((4R,6S)-4,6-
dihydroxycyclohex-2-en-1-
ylidene)acetamide
Purshianin
(E)-2-((4R,6S)-4,6-
dihydroxycyclohex-2-en-1-
ylidene)acetic acid
(E)-2-((4S,5R,6R)-4,5,6-
trihydroxycyclohex-2-en-1-
ylidene)acetamide
Lithospermoside
(E)-2-((4S,5R,6R)-4,5,6-
trihydroxycyclohex-2-en-1-
ylidene)acetic acid
Each of these molecules alone would not cross the cell membrane of the cancer cell. Only those related to
carbohydrate and fulfilling the conditions of item 4. they will block and / or permanently damage her normal
physiology. The use of AAF (from Table 6) directly for treatment will lead to severe toxic reactions and
allergic responses of the body.
By themselves, these compounds or their homologues are still used in conservative chemotherapy
[44÷46]. Glycosides such as Rehmapicroside, Loganic acid, HMBOA D-glucoside, Glucose beta-1,3-
isofagamine, Vanillyl beta-D-glucopyranoside and others. Although they contain AAF of the proposed type,
they would not cross the cell membrane of the cancer cell. They do not fulfill the condition of §5., in the part
of the amido derivative which is to be hydrolyzed by a transitional complex with a carboxylic acid.
The relative inertness of the glycosidic bond (in vivo) also allows the use of different amide-carboxyl
glycosides simultaneously. This is also observed in nature with regard to the distribution of nitrile glycosides
- they are often more than one representative in one plant. Thus, different AAFs can be injected
simultaneously, at different concentrations and at different times, in order to closely differentiate the different
types of cancers, through the synergistic action of the controlled toxicity itself inside the "attacked" cell.
Natural nitrile glycosides would not cross the cancer cell membrane. They decompose to HCN-acid,
phenyl methanol and carbohydrate. They do NOT have anticancer activity due to their inability to reach the
target unchanged. These compounds, in their natural form, are extremely toxic to the human body. Applying
them is not a cure, even at a higher concentration, they do more than they can help. We have theoretically
derived dozens of their modified forms, but their amides and their carboxylic acids are the most promising for
their introduction into conservative oncology. The fact is that the cancer cell itself tries to counteract it in a
fairly certain way.
7. Conclusions
1) The amide derivatives of nitrile glycosides exhibit anti-cancer activity;
2) the cancer cell probably seeks to displace hydrolysis of these derivatives in a direction that would not
pass through its cell membrane;
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3) the amide-carboxyl derivatives of nitrile glycosides could deliver extremely toxic compounds within
the cancer cell itself and thus block and / or permanently damage its normal physiology.
ETHICS APPROVAL AND CONSENT TO PARTICIPATE
Not applicable.
HUMAN AND ANIMAL RIGHTS
No Animals/Humans were used for studies that are the basis of this research.
CONSENT FOR PUBLICATION
Not applicable.
AVAILABILITY OF DATA AND MATERIALS
The authors confirm that the data supporting the findings of this study are available within the article.
FUNDING
None.
CONFLICT OF INTEREST
The authors confirm that this article content has no conflict of interest.
ACKNOWLEDGEMENTS
Declared none.
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