Chapter 1.

Introduction
1.1. Description of the ailment

The discovery long ago has evidence that phytochemicals the naturally occurring compounds
found in plants, hold promise as antioxidants, antidiabetic agents and anticancer activity
including screen various other types of health benefits because many plants also contain
chemicals that are processed results from those biochemical by often help prevent cell damage
within our body. Alkaloids, phenolic acids, flavonoids, terpenoids, and saponins are bioactive
compounds that show extensive biological activity. These processes underlie the remedial
properties of these compounds [1]. Since immemorial, herbal medicine and its pure active
ingredients have been used as dependable medicines. Today, there is an increasing focus on
using natural remedies like crude plant extracts or their bioactive compounds to treat diseases.
Modern medicine has gained significant achievements in treating various ailments. Plants are
the primary nutrient source and fulfill basic requirements such as clothing, fertilizers, shelter,
scents and flavors and significantly impact the healthcare system to prevent various diseases.
At the start of human civilization, ayurveda emphasized the profound relationship between
humans and medicinal herbs [2, 3]. Due to advancements in research, plant-derived
phytoconstituents such as pyridoxal phosphate, ascorbic acid, pregnenolone, flavonoids,
hydroxyl-rich phenolic compounds, sugars, xanthones, etc. are efficient in playing prominent
roles in tackling persistent illnesses such as insulin resistance, oxidative stress-related
malignancies, inflammatory tumors, and viral infections, in addition to investigating the
potential of using drugs with antioxidant that anti-diabetic, and cancer-fighting properties [4-
6]. Ayurveda is among the most ancient medicinal systems that offer countless opportunities
to develop natural medicine and herbal medications currently resurgent in popularity in
developing and industrialized nations due to minimal side effects [7]. The folk healing
properties of these medicinal natural plants have made traditional botanicals a medication
discovery in the last few years and widespread usage of these novel pharmaceuticals to treat
various ailments [8, 9]. They are known for their ability to scavenge free radicals, constituting
a group of antioxidants referred to as phytochemicals. It aids in shielding cells from oxidative
stress, which is harm caused to cells by free radicals. These properties are essential for
managing cellular damage from reactive oxygen species (ROS). Oxidative stress occurs when
there is an imbalance between the generation of free radicals (reactive oxygen species) and the
body's capacity to neutralize these reactive molecules using antioxidants [10]. An
overabundance of ROS can trigger cellular damage, leading to aging and a range of disorders,

1
including tumors and heart disease. In this respect, phytochemicals like flavonoids and
phenolic acids have efficiently provided chelation of metal ions and free radical scavenging,
serving as well as regulators of endogenous defense of antioxidants like glutathione [11].
These systems contribute to the preservation of cellular integrity, reducing the possibility of
diseases associated with redox imbalance. Phytochemicals may have proven potential in
managing diabetes mellitus (DM), a type of metabolic disorder, with chronically elevated blood
glucose levels caused by either low sensitivity to insulin or an absolute lack of insulin
production. Different phytochemicals can enhance glucose homeostasis through several
mechanisms. For instance, flavonoids can increase insulin secretion and sensitivity and, thus,
reduce carbohydrate-degrading enzymes including alpha-amylase and alpha-glucosidase, in
addition to changing the uptake of glucose by cells [12]. Alkaloids like berberine cause the
improvement of insulin signaling pathways and the decrease in liver gluconeogenesis. Since
phytochemicals have a broad range of effects, they greatly assist in the research and
development of novel diabetes medications [13]. The anticancer properties of phytochemicals
are an area of huge research interest. A wide range of mechanisms, furthermore, is linked to
the anticancer activities of these chemicals including induction of apoptosis, cell growth
inhibition, and metastasis inhibition [14]. For example, polyphenols can also influence
signaling pathways overseeing cell cycle control, apoptosis, and angiogenesis [15]. Due to the
potency of modulation of several molecular targets and pathways, phytocompounds are
proving to be highly attractive candidates in the development of a natural health product and
medicines [16].

1.2. Importance of Phytocompounds (PC)

The Critical Importance of Phytonutrients: The Simple Factor of Plant Health and Human
Well-being: Phytochemicals, or Phytocompounds, are bioactive compounds naturally existing
in plants. Phytochemicals are the secondary metabolites of plants and exist mostly as non-
nutrient chemicals. They can be found in differing plant parts, such as leaves, roots, wood,
rhizomes, flowers, bark, fruits, and seeds [17]. The huge interest in these compounds arises
from their potential health benefits, particularly antioxidant, antidiabetic, and anticancer
properties. PCs are important and effective in various areas, such as disease prevention,
therapeutics, and general health and well-being. Phytochemicals are a broad variety of non-
nutrient compounds produced by plants. They have formed the intrinsic defense system of
plants, protecting them from the adverse effects of ultraviolet radiation, various insects,
bacteria, and fungi [18]. For instance, the bright colors that are seen in vegetables and fruits

2
often result from anthocyanins, a phytochemical known to mainly function in protecting plants
from the harsh effects of the sun [19]. Similarly, capsaicin (8-methyl-N-vanillyl-6-
nonenamide), the agent that gives chili peppers their heat, wards off herbivores [20]. Specific
phytochemicals act as innate insecticides and fungicides, repelling herbivores and pathogens,
while others are quite important in plant growth and development, mainly in hormone signaling
and nutrient uptake [21]. Different phytochemicals, including phenolics and flavonoids, have
been shown to possess very strong antioxidant properties. They can endure the harmful effects
of free radicals, which are volatile chemicals that cause cellular damage. Undoubtedly, these
volatile chemicals are responsible for the development of long-lasting illnesses that include
tumors, cardiovascular problems, and cognitive difficulties [22].

Although these PCs are majorly beneficial to the plant, there is a wide array of possible health
advantages that might be gained by humans who consume them. Many phytochemicals have
antioxidant activity, therefore being able to scavenge free radicals in the body that can harm
cells and lead to chronic disease. Allicin (diallylthiosulfinate) is found in garlic, while catechins
are found abundantly in green tea [23]. This could, in addition, reduce inflammation, which is
a major factor in almost all diseases. Some examples of phytochemicals known to have anti-
inflammatory and anticancerous properties such as curcumin, a compound found in turmeric
and resveratrol, available in red grapes and peanuts [24]. On the other hand, it is very well
known that these phytochemicals exhibit therapeutic properties, that they provide nutrition for
normal cell health and maintenance, they enhance the immune system, fight disease-causing
organisms, inhibit carcinogens, and exert antioxidant activities [25]. In this concern,
phytocompounds have been documented to exhibit very promising antidiabetic activities that
make them useful in the management and prevention of diabetes. Alkaloids have been reported
to enhance insulin sensitivity, improve glucose absorption, and control carbohydrate
metabolism, similar to terpenoids and saponins [26].

PCs provide a natural and effective means of controlling diabetes and mitigating its effects by
regulating blood sugar levels and enhancing insulin action. Phytocompounds have exerted
quite considerable anti-cancer activities, hence offering hope in this fight against this
devastating disease. Phytochemicals can slow the growth of cancer cells and trigger
programmed cell death, and prevent invasion processes of cancer cells from one part of the
body to another i.e., metastasis condition [27]. For instance, it has been shown that curcumin
from turmeric or curcuma, resveratrol from grapes, and EGCG (epigallocatechin gallate) from
green tea display potent anticancer activity. Phytocompounds hence provide a multi-

3
dimensional approach toward chemoprevention and chemotherapy for cancer because their
action is on various pathways of carcinogenesis [28].

Plants-derived phytocompounds offer a plethora of new bioactive compounds, some of which

have been evolved for major medicinal agents available in the market today. The worldwide
annual market value of plant-based goods is now expected to reach 77.8 billion US dollars in
2025, which includes both traditional products and innovations. Hence, both academia and
industry have focused their concerns on the direction of medication advancement from
medicinal plants [29]. Integrating the traditional knowledge and implementation of existing
scientific tools could lead to a new way for natural product-based drug discovery. This influx
of high throughput screening, high-resolution screening and hyphenated chromatographic
methods that include (e.g. GC-MS, LC-NMR-MS, LC-MS, etc.) higher magnetic field strength
NMR as well entries into the arena by capillary-distance types have significantly sped up
natural product-based drug discovery efforts [30].

1.3. Oxidative Stress

Free radicals, typically reactive nitrogen and oxygen species (RNS/ROS), are principal
producers of stress. Free radical chemistry: Oxidative stress can either be endogenous or
exogenously induced [31]. Reactive oxygen species (ROS) including hydrogen peroxide,
superoxide radicals and singlet oxygen are generated continuously through the reaction of
oxidative metabolism involving xenobiotics compounds foreign to an organism for example
some drugs or environmental stress conditions which can also enhance the generation of these
reactive oxidant forms [32, 33]. The onset of oxidative stress within the body results from the
disparity between ROS generation and antioxidant defenses [34]. This imbalance leads to age-
related conditions that lead to cell or tissue injury and contributes to many disorders such as
cancer, diabetes, persistent kidney disease, neurodegenerative diseases, long-term obstructive
lung condition, cardiovascular diseases (CVDs), frailty, sarcopenia, and atherosclerosis. As
previously discussed, free radicals are endogenously and exogenously generated. Endogenous
free radical production includes infection, inflammation, excessive exercise, aging, cancer,
mental stress, ischemia, and immune cell activation. Due to the presence of heavy metals such
as Arsenic (Ar), Mercury (Hg), Iron (Fe), Cadmium (Cd), and Lead (Pb) some drugs like
bleomycin, tacrolimus, cyclosporine, and gentamycin, cigarette smoke, radiation, alcohols,
cooking pattern (fat, used oil, and smoked meat) lead to the generation of exogenous free
radicals [35, 36]. Chronic renal diseases are also linked to elevated ROS generation and

4
depletion in antioxidant production leading to problems such as anemia, hypertension,
inflammation, and atherosclerosis [37]. Obesity-related diseases encompass type 2 diabetes,
hypertension, dyslipidemia, stroke, coronary artery disease, and non-alcoholic fatty liver
disease, all of which are associated with the generation of oxidative stress [38]. Excessive
oxidative stress production disrupts the functioning of the neurological system onset of
“amyotrophic lateral sclerosis”, “Huntington's disease”, “Alzheimer’s disease”, epilepsy,
depression, and “Parkinson's disease” [39].

Phyto-antioxidants in various medicinal plants and foods play a pivotal role in curing chronic
diseases generated by oxidative stress and act as an antioxidant by neutralizing ROS and
safeguarding cells. The mechanism of action includes boosting the activity of enzymes such as
superoxide dismutase (SOD) and catalase (CAT), as well as non-enzymatic actions involving
bioactive phenolic and amine substances [40, 41]. From ancient times natural products, and
ingredients have created new medications for medical approaches. Plant-derived
phytocompounds are classified into three primary sectors based on their synthetic pathways.
Nitrogen (-N) containing chemical entities include cyanogenic glycosides, glucosinolates, and
alkaloids; Phenolic compounds like terpenes, flavonoids, and phenylpropanoids. It was found
that dietary phytocompounds have a broader therapeutic effect as they influence different
biological pathways that are associated with disease prevention and health [42, 43]. Natural
bioactive compounds such as quercetin, resveratrol, curcumin, etc., can counteract oxidative
stress-related diseases. On the molecular level, these phytoconstituents activate the Nrf2
signaling pathway that protects against neuronal apoptosis, oxidative stress, and
neuroinflammation conditions like Traumatic brain injury (TBI) [44]. In such cases, the
phytocompounds offer therapeutic potential to target health issues related to oxidative stress
within the body and reduce ROS levels. The toxicants in the environmental system can alter
different biological processes and may also target specific organs. The development of ROS
generation is common due to ecological toxicant pollutants. Still, it varies with the chemical
nature of the toxicant, amount of exposure, targeted route of exposure, time duration of
exposure, and so on. The best way environmental toxicants can induce ROS generation inside
the human body is through inhibiting the antioxidant defense system, dysfunctioning
mitochondria, starting the Fenton reaction, and initiating the production of metabolites and
chemicals that are harmful to the body system [45-47]. Besides, several processes are involved
in the production of ROS such as infection, inflammation, mechanical, and chemical stressors,
exposure to UV rays, and ionizing radiations. ROS act as signaling molecules at basal levels to

5
induce cell proliferation, survival, immunological responses, death, motility, differentiation,
and stress-responsive pathways [48]. Such comprehensive knowledge of oxidative stress can
have a substantial impact; hence, it will have an important outcome on the entire health process
and the development of diseases. Consequently, molecules that arise from oxidation of
proteins, lipids, and DNA are often used as a means to assess the level of oxidative damage,
rather than directly measuring reactive species. Such compounds that arise from oxidative
damage are often collectively referred to as biomarkers [49].

Scientists now agree that the oxidative stress theory is the leading cause of aging and the
problems associated with it. The development of age-related illnesses associated with oxidative
stress and free radicals including diabetes, cancer, inflammatory diseases, cardiovascular
diseases, hypertension, liver, renal injury, neurodegenerative diseases, dementia, osteoporosis,
atherosclerosis, vascular disorders, and metabolic syndromes is significantly influenced by
oxidative stress [50, 51]. The organism has several built-in defense mechanisms, antioxidants,
against a dangerous oxidative environment. These defenses include classical antioxidant
enzymes such as “catalase”, “glutathione peroxidase”, “superoxide dismutase” (SOD), or
“non-enzymatic ROS-scavengers” like “vitamin C” (ascorbic acid), uric acid, vitamin E and β-
carotene [52]. These free radicals not only cause harm in and of themselves, but they also have
the potential to trigger a wide complex network of intracellular stress response pathways, which
in turn cause damage to those cells. Nf-κb, p38 MAPK, hexosamine, JNK/SAPK pathways,
protein kinase C (PKC), AGE/RAGE interactions, and sorbitol production are components that
fall under this category [53].

1.4. Diabetes mellitus (DM)

The history of recorded diabetic patients goes back to the days when Vedas were written. This
was called madhu meha, where "madhu" means honey and "meha'' meant the process of
eliminating through urine. DM, the disease designation used today, basically means that too.
The root of the term diabetics comes from Latin which is “diabētēs” mean passing through.
After all, the ending "mellitus" is derived from Latin for "honey urine” [54]. Rising nations
like India have witnessed a big change in lifestyle patterns & diet habits over the last few
decades. As a result, the aforementioned have changed the state of health and disease. The
prevalence of metabolic diseases, exemplified by diabetes mellitus and hypertension has soared
to epidemic levels India is becoming the world capital for diabetes which again tells the whole
story. Today, 74 million individuals are living with diabetes in India and that is one of the

6
lowest numbers when taken relative to other diseases. India is a diabetes capital together with
the USA and China, collectively responsible for 14% of the global burden; and nearly 95% of
cases are those of type-2 DM (T2DM). Further, the prevalence of diabetes is showing a rising
trend across all geographical and socioeconomic sections in India. It is projected that by 2045
there will be at least 124 million victims of diabetes [55, 56].

The World Health Organization (WHO) describes hyperglycemia as a chronic condition

resulting from the pancreas's failure to produce sufficient insulin or the body's resistance to its
effects, both of which regulate blood sugar. High Blood sugar, commonly called
hyperglycemia, is one of the common conditions caused by uncontrolled diabetes. This
gradually results in significant harm in several systems of the human body, more specifically
in neurons and blood vessels. Conversely, DM is a condition characterized by metabolic
dysfunction with many diverse etiologies that involve a long-term elevation in blood sugar with
impairments Insulin insufficiency leads to disruptions in glucose, lipid, and protein metabolism
production or both (https://www.who.int/en/news-room/fact-sheets/detail/diabetes accessed on
15.06.2024). Indians have major susceptibility to T2DM at younger ages and lower body
weights in comparison with other populations worldwide. Several interesting and not as well-
researched elements might be able to contribute to this particular and increased risk. First,
Indians show a strong tendency for naturally having a lack of insulin secretion or function and
for hepatic steatosis [57]. The overall prevalence of adult-onset diabetes increased from 2% in
1972 to 9% in 2018 in the Indian population. This increase has cut across all sections of Indian
society, including different age groups, genders, socioeconomic groups, and urban/rural areas.
This increase in diabetes prevalence has coincided with rapid economic development and a rise
in the number of overweight people [58]. Among adults aged 18 years and older, diabetes was
found in 8.5% of people in the year (2014). In 2019, diabetes was responsible for more than a
million deaths as the direct cause and in most cases, it is an underlying or leading factor of
death among people aged 20-70 years. Diabetes also accounted for 460,000 kidney disease
deaths; raised blood glucose causes around one in every four cardiovascular deaths [59].

1.4.1. Types of diabetes

There are two primary types of diabetes that are of utmost significance: diabetes mellitus and
diabetes insipidus. Given the persistence and otherwise incurability of DM, we tend to think of
diabetes as diabetes mellitus when referring to diabetes.

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1.4.1.1. Type-1 Diabetes mellitus (T1DM)

T1DM, also known as insulin-dependent diabetes mellitus, is a medical disorder characterized

by the absence of endogenous insulin production in the body. Typically starting in childhood,
this kind of diabetes is marked by the need for daily insulin injections and is not preventable
[60]. The symptoms of this pathology manifest in large quantities of urine production, polyuria,
excessive and intense thirst, polydipsia, considerable hunger, significant weight loss, changes
in vision, and continuous tiredness. All of these symptoms can appear at one time [61].

1.4.1.2. Type-2 Diabetes mellitus (T2DM)

Type-II diabetes, also known as diabetes mellitus without insulin dependence or adult-onset
diabetes, is a case whereby the body cannot use insulin effectively. T2DM is the highest
prevalent type of diabetes in the world and mainly results from excessive body weight and lack
of physical exercise [62]. Symptoms are just similar to T1DM. This kind of diabetes, once
limited to adults, is becoming more prevalent among young individuals.

1.4.1.3. Gestational diabetes mellitus (GDM)

GDM is a medical disorder wherein, due to a raised glucose level, women are found to have
difficulty in processing the glucose in their bodies. In about 5-7% of pregnancies, it is noted
that GDM causes a wide range of effects on the mother and the fetus, such as abnormal fetal
development and a heightened susceptibility to developing T2DM in the future [63]. In most
cases, gestational diabetes is diagnosed by prenatal screening and not due to reported
symptoms.

1.4.1.4. Impaired glucose tolerance (IGT) and impaired fasting glycemia (IFG)

There are states in between having normal blood sugar and having diabetes. If one is diagnosed
as having either IGT or IFG, then the odds of eventually developing T2DM are much higher
than normally expected, but not an inevitability. Research has indicated that the risk increases
for IGT by 35% in overweight subjects and by 77% in those who are obese. Moreover, it
increases by 122% for IFG [64].

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1.4.2. Standards for diabetes diagnosis

The criteria for assigning an individual as diabetic can be evaluated through blood glucose level
(mg/dl), mentioned in Table 1 and the glycated haemoglobin (HbA1c) value described in Table
2. [65].

Table 1. Presents WHO Recommendation for declaring a person as diabetic

S. Condition of individual Blood glucose level (mg/dl)

No.
1. The random blood sugar level is indicative of diabetes > 200 mg/dl

2. Fasting Glucose Level > 126 mg/dl

Table 2. Confirmation and monitoring for diabetes are done via the HbA1c test

S. Condition of individual HbA1c level

No.
1. Normal value 4%-5.6%
2. Diabetic risk 5.7%-6.4%
3. Diabetic > 6.5% (or above)

1.4.3. Indicators of diabetes mellitus

The most common symptoms of diabetes include discomfort, heartburn, dyspnea, weariness,
edema, dysphasia, cognitive impairment, asthenia, and somnolence among other experiences
that typify type 2 diabetes. More specifically, frequent urination, excessive thirst, intense
hunger, weight loss, exhaustion, irritability, and hazy vision are some of the characteristic signs
among these individuals, those with both type 1 and type 2 diabetes experience fluctuating
blood sugar levels [66] are shown in Figure 1.

9
Figure 1. Symptoms of Diabetes mellitus.

1.4.4. Downstream consequences of diabetes

Diabetes mellitus can also lead to many problems due to the high incidence of hyperglycemia.
Ultimately, the degree of blood sugar regulation determines the probability of acute or chronic
problems occurring.

1.4.4.1. Acute Complications:

1.4.4.1.1. Hypoglycemia

Hypoglycemia is usually one of the most prevalent diabetes symptoms, especially among
people living with T1DM and T2DM, as evidenced by many published research articles [67].

1.4.4.1.2. Ketoacidosis

Diabetic ketoacidosis (DKA) is a serious complication that ensues from diabetes mellitus,
characterized by high sugar blood levels, the presence of ketones, and an imbalance in the

10
body's acid-base equilibrium. DKA is a complication most times initiated by causes that
include infections, lack of medication adherence, and metabolic stress circumstances [68, 69].

1.4.4.1.3. Non-Ketogenic Hyperosmolar Syndrome

HHS, which stands for Hyperosmolar Non-Ketogenic Syndrome is a severe complication of

diabetes mellitus that occurs to those who have T2DM. It is distinguished by severe glucosuria
and hyperglycemia but without typical symptoms of ketoacidosis. Less common is
hyperosmolar hyperglycaemic state (HHS) previously called non-ketotic hyperglycaemic
coma; these patients present with alterations in mental function, high total body water losses
and potentially focal neurological deficits due to the result of profound dehydration, which
carries a mortality of up to 20% [70, 71].

1.4.4.2. Chronic complications:

Interpreting illness in long-standing diabetes patients is very complex, characterized by

recurrent increases in blood glucose levels. Two main participants, glycosylated proteins and
intracellular accumulation of sorbitol, are responsible for developing most of the chronic
problems of diabetes [72]. Chronic complications of diabetes: These are a wide array of long-
term problems in health that may influence the quality of life and, eventually, mortality of the
patients associated with diabetes. Literature available has shown that most people living with
diabetes often experience long-term complications that include cardiovascular problems,
endocrine/metabolic problems, and neurological problems. Of the three, 77% show a high
prevalence rate for such problems [73].

1.4.4.2.1. Glycosylation

Nonenzymatic glycosylation involves a no-enzyme attachment of sugars to proteins, nucleic

acids, and lipids. The attachment gives rise to early glycosylation products, otherwise described
as Amadori products. The EGPs are oxidizable to give dicarbonyls that will then constitute the
irreversibly formed and very reactive AGEs linked structures. AGEs formation is an
irreversible process and continues to build up in various tissues of the body, thus ensuring
further binding to or alteration of proteins both inside and outside cells and also the formation
of reactive oxygen radicals (ROS) [74-76]. This can have a variety of harmful effects, including
inactivation of enzymes, reduction or prevention of the binding to regulatory molecules, cross-

11
linking or glycosylation of proteins, entrapment of soluble proteins by the glycosylated
component of the extracellular matrix, reduced susceptibility toward proteolysis, abnormalities
in the function of nucleic acids, altered recognition of macromolecules, and increased
immunogenicity [77].

1.4.4.2.2. Atherosclerosis

Compared with those without diabetes, the risk is much higher two to threefold greater in
people with T2DM. This is not to take the connection between these factors that have been
found through many investigations. Atherosclerosis development in diabetic vessels, mainly
due to inflammation and other factors originating from hyperglycemia, is faster than that of
people with normal glucose levels; studies have found a higher rate of macrovascular events
(e.g., heart attacks or strokes) among those who suffer diabetes compared to those without the
disease [78].

1.4.4.2.3. Diabetic retinopathy

This eye disease is highly hazardous and potentially blindness-causing. Type I diabetes
individuals are at the same risk of developing retinopathy as those who have type II diabetes.
A common problem that can happen with diabetes is diabetic retinopathy (DR) and consists of
continuous damage to blood vessels in the retina, causing visual loss that can progress through
blindness. DR affects around 90 million individuals globally, which means it's very common.
It is a common cause of vision loss in people of working age [79, 80].

1.4.4.2.4. Diabetic nephropathy

This condition is common among individuals with diabetes mellitus and is a major cause of
mortality in this group. Diabetes can lead to a serious kidney complication called diabetic
nephropathy, which may advance to kidney failure in its final stages. It is marked by
microalbuminuria, which ends with proteinuria. It is influenced by hyperglycemia, proteins
altered by high blood glucose levels, reactive oxygen species, growth factors, cytokines, and
the renin-angiotensin system [81, 82].

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1.4.4.2.5. Diabetic foot ulcers

Probably the most common and troubling complication of diabetes is Diabetic foot ulcers
(DFUs), affecting as many as 15-25% in a lifetime. These occur due to such factors as poor
control of blood sugar, damage to the nerves, and reduced circulation to the extremities. This
often initiates chronic foot ulcers, which can worsen and eventually necessitate the amputation
of a lower leg [83, 84].

1.4.4.2.6. Oxidative Stress

One of the pervasive reasons for diabetes mellitus is oxidative stress, which induces issues like
neuropathy; retinopathy and nephropathy among others by interacting with numerous
biomarkers as well as enzymatic systems. The biomarkers: malondialdehyde (MDA),
superoxide dismutase (SOD), glutathione (GSH) and catalase (CAT) can be used to understand
the oxidative stress occurrence in diabetes. Elevated levels of these markers reflect the presence
of oxidative stress that can be used as a premature predictive mark for diabetic complications
[85, 86].

1.4.5. Possible contributing factors to diabetes

Cause of diabetes is one the most common and chronic illnesses people are facing all around
the world. Some environmental agents too like water pollution, wrong food habits and irregular
physical activities lead to basic disturbances in the level of hormones forming an empire for
breeding this illness environment by stress and exposure including viruses also play a role.
Diabetes is essentially a multifactorial condition that develops as the result of genetic
predisposition, lifestyle choices and hormonal factors. Diabetes research that identifies the
strength of risk involved in getting diabetes is that it has a direct relationship between variables
like age, FHx DM: Family history of Diabetes mellitus, BMI:Body mass index and also early
marriage, delayed delivery & using exogenous hormones, etc., and more are listed in Figure 2.
Impaired insulin sensitivity and disrupted beta-cell activity, usually related to the metabolic
setting induced by Western lifestyle in genetically predisposed individuals are typical features
of non-insulin-dependent diabetes mellitus [87, 88].

13
Figure 2. A schematic diagram of potential contributors to hyperglycemia in diabetes mellitus.

1.4.6. Insulin's mechanism in glucose homeostasis

Insulin-mediated glucose transport is an important mechanism in tissues that use or store

glucose. Insulin activates the intracellular enzyme glucokinase, resulting in more rapid glucose
phosphorylation and an increased rate of glucose oxidation [89]. It activates glycogen
synthetase, stimulating the production and retention of glucose as glycogen in both muscles
and the liver. This also serves to attenuate the glycogenolysis inhibitory effect on blocking of
phosphorylase. Insulin diverts glucose to be used preferentially as an energy source while
sparing fat and protein from oxidation [90]. An anabolic counter-regulatory hormone, one that
antagonistic the effectiveness of catabolic hormones, including adrenaline, glucagon and
growth hormone. And it disrupts gluconeogenesis. Glucose absorption is crucial in muscle,
adipose tissue, and liver, while it suppresses lipolysis [91]. In the absence of insulin, there will
be continuous accumulation of glucose in the blood and liver. The manufactured amino acids
are used in gluconeogenesis while the fatty acids are changed to ketone bodies; the production
of the counter-regulatory hormones within the body increases [92, 93].

14
1.4.7. Treatment of diabetes

There is currently no pharmaceutical intervention that can fully eradicate diabetes. However
various strategies are present to solve this global problem. Like as Allopathic medication,
herbal or ayurvedic formulations, yogic or exercise, and lifestyle changes. A traditional
Chinese medicine for the treatment of diabetes consists of Dioscorea, astragalus, Chinese
atractylodes, salvia, figwort, rehmannia, pueraria, coptis, lycium bark, mume, coptis, and poria.
These herbs are combined with a broad range of health benefits [94]. Standard western
medicine is needed to manage diabetes, T2DM in particular. In the case of T2DM, patients
often move from oral hypoglycemic drugs to insulin injections because their blood sugar levels
are poorly controlled and they experience gastrointestinal disturbances. Details While
allopathic diabetes treatments like insulin sensitizers, or alpha-glucosidase inhibitors are the
usual prescriptions given in primary care for T2DM; long-term use of these drugs can be
counter-productive. Data support the exploration of natural product agents to provide
additional options for patients [95, 96]. The most important allopathic antidiabetic drugs
include biguanides, sulfonylureas, DPP-4 inhibitors, GLP-1 agonists, SGLT-2 inhibitors,
thiazolidinediones, and the different types of insulin [97]. These allopathic antidiabetic drugs
have some side effects, such as kidney impairments, malabsorption of nutrients, excess gas,
diarrhea, and abdominal swelling. Therefore, there is a need for safer alternatives, and herbal
preparation is one among them [98].

Various herbs and dosage forms have well-documented potential in the treatment of diabetes,
and Ayurvedic books reaching back to ancient times bring to light several formulations for the
control of “Prameha” (Diabetes mellitus) [99]. Ayurvedic herbs screened for antidiabetic drug
potential are quite promising in the treatment of DM. Herbal formulations contain
phytochemicals like polyphenolic compounds, alkaloids, glycosides, flavonoids, and
terpenoids. The endocrine potential for antidiabetic action is mediated by the stimulated release
of insulin from the pancreas while decreasing glucose synthesis in the liver-bound fraction
[100]. Plants like Trigonella foenum-graecum, Aloe vera, and Andrographis paniculata have
been known for their potential to decrease blood glucose through various mechanisms such as
GLP-1 release and inhibition of enzymes [101]. Herbal drugs are a great option to supplement
traditional methods of diabetes control because of their low cost, lack of side effects, and safety
profiles. Different studies have proved the potential of Ayurvedic formulations as antidiabetic
drugs. For instance, it is documented that polyherbal formulations such as “Dolabi” are useful

15
in lowering blood glucose in subjects with hyperglycemia. In addition, “Vidangalihouham”
exhibited significant antihyperglycaemic activity in animal models of diabetes characterized
by reduced levels of fasting blood glucose with improved biomarkers [102, 103].

This practice of yoga in balancing harmony in the physical, mental, and emotional self was
followed by people in India for ages spanning over five thousand years. T2DM is one of the
several lifestyle disorders which can be helped through the regular practice of yoga. These
protective benefits of yoga on diabetes result from the activation of the immune system and
psychno-neuro-endocrine processes [104]. There are numerous studies that have documented
significant antidiabetic advantages of yogic practices. Yoga supports diabetics by improving
pancreatic function and reducing mental stress, promoting unity of mind, body and emotions
[105]. Further scientific research also states that by doing regular yoga, pranayama &
meditation one can reduce their medicine dosage with some conditions, for example, T2DM.
It demonstrates the potential of yogic practices as affordable and safe therapies with minimal
or no adverse effects [106]. In general, yoga activities have been suggested as a useful
extension of conventional diabetes mellitus therapies that offer cost-effective advantages in the
treatment of this illness.

For lowering metabolic control lifestyle therapies are important and they can help in retarding
the development of diabetes in an individual with poor glucose tolerance or prediabetes [107].
They are linked to reduced indices of plasma glucose while fasting and 2-h blood sugar levels,
which in turn is also associated with decreased diabetes risk. Also, significant lifestyle changes
have shown a 20% relative risk reduction in diabetes. This makes them an effective protective
strategy, along with metformin also interested in the prevention of prediabetes [108]. This
reduces the risk of diabetes and its complications through regular physical activity, a balanced
diet, and a small reduction in body weight. Such behavioural modification also helped
normalize blood sugar and had a significant effect on regulating hyperglycemia [109].

1.5. Cancer

Carcinoma is a diverse collection of disorders defined by atypical cell proliferation and

differentiation. Types include sarcoma, carcinoma, lymphoma and leukaemia as well as
myeloma each in a kind of area [110]. The process of developing cancer involves the
accumulation of genetic alterations that transform normal cells into malignant ones. It is well
known that these cells can initiate tumors, benign or malignant according to their potential for

16
invasion and metastasis [111]. Risk factors that may contribute to the initiation of cancer
include a range of lifestyle choices such as tobacco smoking, eating unhealthy food and not
exercising enough; infections from pathogens including human papillomavirus (HPV) and
hepatitis [112]. Treatment options for cancer are plentiful and depend on the exact type of
cancer as well as its stage. These treatments such as encompass surgery, chemotherapy
medication radiation therapy [113]. These were, and are, highly relevant categories to WHO
(World Health Organization) for the diagnosis and treatment of cancer. Classification issues,
firstly, about the histological properties of tumors, and secondly, about their molecular and
genetic characteristics were key and core issues. This way, tumors are more precisely classified
and then also further help guide therapeutic decisions [114]. It is WHO, therefore, that has
promoted policies generated from centers of governing bodies within it by emphasizing the
importance of cooperation between international organizations and oncology specialists, which
keeps cancer control at the forefront of the worldwide medical agenda. For example, the 70th
World Health Assembly Resolution "Cancer Prevention and Control in the Context of an
Integrated Approach" was adopted in 2017 by 194 Member States of the WHO to tackle the
growing global disparities in cancer-related outcomes[115].

1.5.1. Cervical Cancer

Cancer is a significant killer and incapacitating disease condition worldwide. Regarding to

cancer deaths, cervical cancer remains the fourth most lethal form in women, accounting for
604,127 cases, with a rate of 6.5 percent, causing 341,831 deaths at a rate of 7.7 percent in
2020 [116]. The percentage of cervical cancer in India is almost 6-29% of the total cases. In
this regard, advanced-stage cervical cancer shows a very poor prognosis, and it is difficult to
identify potential prognostic markers [117, 118]. The illness usually primarily originates in the
area surrounding the cervix at the junction of the endocervical and exocervical epithelium
known as the transformation zone. In this scenario, the involvement of HPV testing in the
screening program becomes of prime importance in diagnosing the disease at an early stage
and managing it properly [119]. The most prevalent postcoital bleeding is an early indication
of cervical cancer, metrorrhagia, or menorrhagia leading to chronic anemia, vaginal discharge,
dyspareunia, low backache, intermenstrual bleeding, chronic back discomfort, pain in the
pelvic area, post-coital bleeding, haste in urination, unexpected loss of weight, and significant
edema in one or both legs [120, 121].

17
HPV, or human papillomavirus, is a common risk factor for cervical cancer, engaging in
intimate relationships at a young age and having several sex partners and many children yet no
HPV vaccination given to the woman or her partner if a man is known as gripping and non-
giving male circumcision community, smoking (> 12 cigarettes a day light up five years at least
one of lung carcinoma case causing chemicals expect you know what family pack results use)
rarely taking oral contraceptives not having been diagnosed with chlamydia trachoma test
reading few Memphis forever watch education maybe so much price bean cars low-cost weight
gain moderate physical activity/ risk friendly behavior increased HPVI infection Nevada cheap
cederos shot gamble strength against other acts urging depressive sex hormones confuse
scandal enhance obesity make eager onset choose embarrassing prepare advancement lady
wages consequences attend meaningful ceremonies. Studies have reported that these factors
are strongly linked to a higher likelihood of developing cervical cancer [122, 123].

The current therapeutic for cervical cancer include surgery, chemotherapy and immunotherapy.
The effect of type, quality and quantity may account for morbidity rates (illness) seen in the
patients. Consequently, natural materials have become increasingly important as more effective
and efficient therapeutic options for combating this type of cancer are urgently needed. Plants
have attracted a lot of interest from humans over time and are popular as they offer some natural
products for human health. Moreover, both scientific and traditional studies have indicated
plants with bioactive compounds named phytochemicals are being investigated for their
promising treatment of diverse diseases and disorders [124]. These are factors related to
socioeconomic, education level achieved, age at first sexual activity and other risky behaviour
relative to history; a few examples would be some aspects related to personal hygiene habits
(Attribute), use of tobacco (Exposure1), or the source for oral contraceptive used in youth;
insufficiencies on diet (Related FactorA); between another plenty more [125]. Moreover, a
panel of plasma proteins such as apoAI, ATIII, apoE, CLU, mTOR and VIL1 displayed 97.6%
sensitivity and specificity for early-stage cervical cancer discrimination. These are crucial
markers for early diagnosis [126]. Machine learning models have been explored in predicting
the early risk of cervical cancer and reached premier rates of accuracy, especially using the
Multi-Layer Perceptron method [127].

The HeLa cell line is a highly established and extensively used human cell line in current
research that has been intentionally dividing indefinitely since they were first pulled from a
young black woman diagnosed with cervical cancer. Cervical cancer is the fourth fastest-

18
growing form of cancer in women and is a significant cause of mortality in females. From here
we can draw the population. A total of 56,9847 newly diagnosed cases were recorded with
about 31,1365 deaths resistance has occurred globally by this disease as studies go for the year
2018 [128].

1.5.2. Prostate Cancer

Prostate cancer is the second most prevalent type of cancer, surpassed only by skin cancer
affecting the prostate gland, frequently diagnosed oncosis in males and represents the most
prominent cause of male mortality from all cancers worldwide. Prostate cancer (PCa) may not
show any symptoms in the early stages and it often grows slowly, possibly requiring
observation only. GLOBOCAN 2018 estimates showed that prostate cancer was the reason for
as many as 13,32,985 new cases globally in developed countries also had a higher incidence
rate [129]. Prostate cancer is a common and malignant creates harm in males internationally,
triggering considerable morbidity along with fatality Multiple elements, such as age, ethnic
background and family history contribute to how the illness develops. The finding also
underscores the importance of early detection and addressing the risk factors such as
maintaining a healthy weight or eating more fruits and vegetables as well [130]. Different
treatments are available which include surgery, radiotherapy, hormonal therapy and
immunotherapy. In the setting of low-grade tumors that are not progressing, active surveillance
may be recommended to preserve quality of life. Improvements in the care of late-stage PCa
have directly translated into better patient survival times and quality of life, highlighting the
evolving landscape for the management of this disease [131].

Phytochemical therapy against prostate cancer garnered significant attention because of its
possible anti-cancer properties and low toxicity compared to standard pharmaceutical drugs.
Many studies have been conducted on the potent activities regarding cell proliferation
inhibition, apoptosis, and angiogenesis exhibition using different bioactive compounds derived
from plants. Different mechanisms are involved in the modulation of phytochemical activity
through epigenetic alterations, comprising histone changes, DNA methylation, and control by
miRNAs and lncRNAs [132, 133]. Androgens and their receptor (AR) are key in the regulation
of prostate cancer's growth or progression. Most patients respond to both surgical and medical
castration, although a proportion of patients relapse following these treatments. It is now clear
from genomic profiling that cancer cells exploit the dysregulated activity of PI3K,
Ras/Raf/MAPKinase and epidermal growth factor receptor (EGFR) signaling pathways to gain
19
a survival advantage. Hence, it may provide therapeutic benefits to target interconnected
upstream development pathways in conjunction with androgen deprivation [134]. Regarding
the potential beneficial effects of phytochemical therapy on prostate cancer, overuse has been
particularly endeared by its suppressive actions in decreasing carcinogenesis and progression
without overt toxicity beyond doses that are widely used as food. Several plant-derived
components, such as resveratrol, tea polyphenols, minerals, and different phytochemicals
including isothiocyanates, lycopene, curcumin, anthocyanins, soy isoflavones, garlic
compounds, luteolin, coumestrol and hesperidin and indoles, have demonstrated potential in
assisting with the management of prostate cancer [135, 136]. These phytochemicals disrupted
major biologic signaling pathways, induced cell cycle arrest, activated apoptosis, inhibited
prostate cancer cells from proliferating, and decreased angiogenesis. Coadministration of these
natural compounds in preclinical studies with hormone or chemotherapeutic agents increased
therapeutic effectiveness and reduced toxicity. Such research would then be the way forward
to opening the avenues for strategies that belong to precision and customized medicine,
improving prostate cancer management [133, 137].

1.6. In Silico Studies

Computational studies with the aid of computers should be made mandatory in discovering
phytochemicals that can have potential differential medicinal applications. These
computational tools will help screen the vaccine or drug candidate molecules against the
ramifications of various diseases like T2DM, antibiotic-resistant infected wounds, and deadly
disorders like diabetes, cancer, and cardiovascular diseases [138, 139]. As drug design moves
toward precision medicine, in silico technology is making it easier and less expensive to tailor
therapies for specific patients. The use of a computational approach is essential in the
advancement of plant compounds-based research and this is important because it provides an
affordable alternative to filter out prospective therapeutic agents from diverse natural chemical
sources. Computational methods, such as molecular docking and quantitative structure-activity
connection studies and ADMET predictions are useful for facilitating the selection of candidate
compounds [140]. Phytochemical protein interaction by utilizing in silico methods can be
pivotal to predict the pharmacological behaviour of phytochemicals and their targets that will
help fasten the drug development process with less emphasis on extensive resultants involving
animal trials for this essential first step before dedicating a large resource [132].
Phytochemicals originating from medicinal plants can target key proteins that partake in the

20
administration of diabetes, e.g., α(alpha)-glucosidase and α(alpha)-amylase; hence they may
play crucial roles in lowering blood sugar [141]. In silico studies of phytoconstituents docking
with different classes of the cysteine-dependent protease family called caspases have been
carried out. Caspases are involved in several biochemical processes such as proliferation,
inflammatory responses, and diversification. They have a vital role in initiating and carrying
out programmed cell death in cancer cells [142, 143]. The caspase family is further grouped
based on the functional contribution. Caspase-2, 8, 9, and 10 activate or initiate apoptosis or
cell death, while caspase-3, 6, and 7 execute cell death [144]. Molecular docking, virtual
screening and bioinformatics databases can offer a variety of information about phytochemicals
for prediction on binding mode with their respective target proteins accelerated drug discovery
process in this field. Computational approaches not only enhance the understanding of
phytochemical properties but also present cost-effective and time-saving solutions to
traditional in vitro and in vivo screening methods, thus extending the capacities for exploration
both on theoretical study and practical usage of plant resources sourced new medicines [145].
The most important practical application is related to anticipating interactions between
phytocompounds this can be done by Computational models such as docking of molecules,
virtual testing, and quantitative structure-activity relationship (QSAR) are utilized. These
models rely on advanced bioinformatics techniques like AutoDock Vina for progressing the
interactions between plant-derived phytochemicals and their biological targets. These
approaches encompass machine learning, network pharmacology and molecular dynamics
simulations both in herb-drug interactions prediction as well as inhibitor identification or
safety/efficacy estimation of phytocompounds [146, 147]. There are many docking software
packages used in computational drug design that predict with high accuracy how ligands will
interact with proteins. The poses of ligands that bind within structures have been explored using
systems such as AutoDock Vina, DOCK 6, Glide, RxDock, UCSF DOCK, PyRx, CBDOCK-
2, etc.

1.7. In Vivo Studies

In vivo studies are conducted for the determination of efficacy, toxicity, and bioavailability for
pharmacological products such as RNAi formulation, nanomedicines, and embolization
chemicals. Indeed, they involve the dosing of live animal models with pharmaceuticals to study
cellular, tissular, and organismic effects [148]. In vivo, mouse tests are a necessity in
understanding the physiology behind obesity and diabetes. These require multi-organ

21
techniques that guarantee little or minimal stress responses to guarantee data integrity during
research procedures [149]. These in vivo models have served as key tools in demonstrating
natural compounds' capabilities in controlling insulin resistance, improving lipid profiles, and
reducing oxidative stress, hence being promising candidates to facilitate the advancement of
novel antidiabetic medications as depicted in Figure 3. The Streptozotocin (STZ) model is very
commonly used in the scientific arena for the induction of diabetes in animal models so that
different substances can be tried on these diabetes treatments. Many studies have been shown
to demonstrate the efficacy of many extracts in the rat model with STZ-induced diabetes [150].
Considering this, the STZ in vivo paradigm is mainly utilized for the induction of diabetes in
animal studies. Various key features include modeling T1DM through the induction of loss of
pancreatic islet β-cells, resulting in insulin deficiency and a corresponding increase in blood
sugar levels. However, STZ can be employed to stimulate the development of a model animal
for Type 2 Diabetes Mellitus (T2DM) and provide a useful tool in investigating its pathogenic
effects on diabetes and exploring possible treatment strategies [151].

Figure 3. In vivo assessment of plant-derived phytochemicals for antidiabetic properties.

1.8. Plant Selection Metrics

Additional wider researches need to be carried out on plants with antidiabetic properties based
on earlier literature and traditional knowledge/practice. There is, however, a criterion for plants

22
being taken on into further research. Several scientists have proposed a set of five criteria as
potential guidelines for prioritizing the species to investigate antidiabetic potential in plants
with research purposes acceleration [152] are as follows:

1) Plant distribution and availability

2) Ethnomedical use for diabetes management

3) In Vivo Safety Profile

4) Demonstrated antihyperglycemic Effects

5) Unidentified blood sugar-lowering compounds

The plant materials under investigation were chosen from a very broad spectrum of sources,
such as medicinal plants that can be cultivated at home, wild species, and even industrial by-
products. It has been noted that various plants of different species were alike in their
morphological features but different from each other in their phytochemical characteristics.
This study aimed at identifying plant extracts and compounds that exhibited antioxidant
activity, antidiabetic activity, and anticancer activity can be visualized in Figure 4 and also
mentioned below:

1) Aqueous (Aq.) extract of Zingiber officinale rhizome (AZOME)

2) Methanolic (MeOH) extract of Allium sativum (MeAS)

3) Aq.+ MeOH + Chloroform extract of Momordica Charantia (MoCtia)

4) Methanolic extract of Trigonella foenum-graecum (MTFG)

5) Methanolic extract of Azardica indica (leaf + bark) (MeAI)

6) Aqueous extract of Cordia myxa stem (ACMS)

7) Policosanol (Polico) compound from rice bran wax (rice mill industry byproduct)

23
Figure 4. Schematic illustration of overall research methodology.

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