Antithrombotic and Anti-Inflammatory Health Promoting Pharmacological Properties of Chalcones and Their Derivatives Against Atherosclerosis and CVD

Abstract

Chalcones, a class of flavonoid compounds, are recognized for their unique biological properties, and especially for their antithrombotic, anti-inflammatory, and antioxidant health-promoting properties against inflammation-related disorders. Chalcones are phytochemicals naturally found in plants, fruits, and vegetables, such as tomatoes, apples, and licorice. Their characteristic chemical structure, which includes two aromatic rings and an α,β-unsaturated carbonyl group, makes them particularly versatile for pharmaceutical use. At the same time, chalcones exhibit strong antioxidant activity by neutralizing free radicals and enhancing endogenous antioxidant defense systems, such as glutathione. Structural modifications have improved their biological activity, leading to important applications in the treatment of atherosclerosis and cardiovascular diseases, cancer, neurodegenerative diseases, and inflammatory disorders. In addition, they have been successfully used in agriculture as natural pesticides and in the food industry as antioxidant additives. This review demonstrates the interdisciplinary importance of chalcones, highlighting the need for further research into their molecular mechanisms of action. A deeper understanding of their properties may open new avenues for the development of innovative drugs and environmentally friendly applications. In this way, chalcones can be a decisive factor in improving human health and environmental sustainability.

1. Introduction

Chalcones, as a subclass of flavonoids, have attracted considerable scientific interest due to their diverse pharmacological activities, including antioxidant, anti-inflammatory, and antifibrotic effects. These properties are not only relevant to atherosclerosis and cardiovascular diseases but also highly pertinent to sclerosis-related disorders. In multiple sclerosis (MS), chalcones have been reported to reduce neuroinflammation, modulate oxidative stress, and exert neuroprotective actions. Similarly, in systemic sclerosis, chalcones may downregulate fibrotic mediators, inhibit fibroblast proliferation, and improve endothelial function, thereby addressing the key pathological features of the disease [1].

The history of chalcones begins with the first synthetic attempts that took place in the 19th century. Stanisław Kostanecki and Josef Tambor, who are known as the pioneers in the synthesis of synthetic chalcones, introduced their chemical name and developed production methods that remain useful even today [1]. Chalcones naturally occur both in their free (aglycone) form and as glycosides. They have been detected in a wide range of plants, fruits, and vegetables, including apples, tomatoes, citrus species, and licorice. Their broad presence in nature has encouraged extensive research, since these compounds play a significant role in the human diet and in various traditional medicinal systems [2].

The antithrombotic and anti-inflammatory actions of chalcones are two of the most interesting aspects of their pharmacological significance. The antithrombotic effects of these compounds are attributed to their capacity to inhibit platelet aggregation and enhance blood circulation, while their anti-inflammatory actions are attributed to the inhibition of inflammatory pathways, such as the inhibition of cyclooxygenase (COX) and lipoxygenase (LOX). Furthermore, chalcones have demonstrated efficacy in modulating reactive oxygen species (ROS) and alleviating oxidative stress, traits that are closely linked to the prevention and management of various chronic diseases [3]. The chemical structure of chalcones is key to their biological properties. They are derivatives of 1,3-diphenyl-2-propen-1-ones, which include two aromatic rings connected via an α,β-unsaturated carbonyl group. This structure makes chalcones extremely versatile, allowing them to undergo various chemical modifications to enhance their pharmacological properties. In particular, the presence of phenolic groups in their rings enhances their action as antioxidants, while variations in substituents can increase biological activity [4].

Chalcones have been found to modulate various molecular mechanisms, notably by preventing the activation of nuclear factor kappa-B (NF-κB) and reducing the production of pro-inflammatory cytokines like IL-6 and TNF-α. These mechanisms are critical for the management of inflammatory diseases, including autoimmune disorders and rheumatoid arthritis. At the same time, the ability of chalcones to interfere with the signaling of the MAPK and PI3K/Akt pathways reinforces their importance in research into treatments for cancer and chronic inflammation. In conclusion, the analysis provided in this paper highlights the interdisciplinary nature of chalcones and their variety of applications [5]. Copper ions appear to serve a key role in controlling inflammatory processes and thrombotic processes, making them candidates for use in pharmaceutical products. In particular, studies focusing on structural modifications of chalcones show that these changes can dramatically improve their biological performance, while providing new opportunities for the development of environmentally friendly agrochemicals and therapies. Extensive research on chalcones has resulted in the identification of numerous synthetic and natural derivatives with specialized properties. These derivatives have been exploited in various fields, such as agriculture (as pesticides and plant growth regulators), pharmaceuticals (for the development of new drugs), and the food industry (as natural additives). At the same time, chalcones have proven useful in addressing diseases, including cancer, cardiovascular disease, and neurodegenerative diseases, by targeting numerous molecular mechanisms. In addition, the use of chalcones as natural antioxidants has been extensively studied. Their capacity to scavenge free radicals and strengthen endogenous antioxidant mechanisms, including glutathione, renders them important for the prevention of degenerative disorders. Furthermore, their ability to reduce oxidative stress in the brain suggests potential therapeutic applications for diseases such as Alzheimer’s and Parkinson’s.

This review aims to deepen our understanding of the antithrombotic and anti-inflammatory properties of chalcones by examining their chemical, biological, and pharmacological aspects. Furthermore, it seeks to highlight the importance of structural modifications of chalcones in enhancing their therapeutic properties. By systematically reviewing the latest research, this paper seeks to offer a thorough overview of the role of chalcones in contemporary pharmacology and their potential for clinical application.

2. Chalcones, Origin, Sources, Structure, and General Uses/Applications

2.1. Origin of Chalcones

The name chalcone is derived from the Greek word chalcos, meaning “copper,” in reference to the yellow–orange coloration characteristic of many chalcone compounds. This etymology reflects their appearance rather than any chemical connection with the metal copper [1]. The first studies on chalcones focused on their synthetic preparation. Stanisław Kostanecki and Josef Tambor, pioneers in flavonoid chemistry, successfully synthesized chalcones in the late 19th century and introduced the term chalcone to describe this class of open-chain flavonoids. Their work established the basic methods of chalcone synthesis, many of which remain in use today [2]. By contrast, naturally occurring chalcones were not isolated until 1910, when plant-derived chalcones were first identified. This marked the beginning of systematic research into chalcones as secondary metabolites with biological roles in plants and, later, as pharmacologically active natural products in humans [6].

As early as the 19th century, several researchers worked on synthesizing chalcones, with Kostanecki and Tambor acknowledged as the pioneers who successfully prepared synthetic chalcones by treating o-acetoxychalcone dibromides with alcoholic alkalis [6]. In fact, Kostanecki and Tambor coined the term chalcone [1], and the word was first introduced in 1899 based on the discovery of mono-oxychalcone [7]. Other widely known chemical names for chalcone include benzyl acetophenone or benzylideneacetophenone [8], phenylstyryl ketone, β-phenyl-acrylophenone, α-phenyl-β-benzoyl-ethylene, etc., and form the central core of biologically active heterocyclic compounds [9]. It was not until 1910 that naturally occurring chalcones were first isolated [8].

2.2. Structural Features

Chalcones are classified within the flavonoid group of phenolic compounds [6]. As open-chain flavonoids, they are derivatives of 1,3-diphenyl-2-propen-1-ones and consist of two aromatic rings connected by an α,β-unsaturated carbonyl system (Figure 1). Due to the double bond in this system, chalcones can exist in two stereoisomeric forms: the trans (E)-isomer, which is the most stable and commonly found in nature, and the less stable cis (Z)-isomer. Both configurations share the same core structural features but differ in the relative orientation of the substituents around the C=C bond [1]. They are based on two aryl units bridged by an α,β-unsaturated carbonyl group [10] and are not only important precursors for synthetic manipulations, but also constitute an important component of natural products [11]. The chalcones in their aromatic rings have a delocalized p-electron order [10]. Specifically, their biological activities are believed to be attributed to the presence of this double (π) bond, in conjunction with the carbonyl property, as the removal of the carbonyl makes them inactive substances [11].

Figure 1. General structures of chalcone alloys [6].

Due to the inclusion of a ketoethylenic CO-CH=CH- structure and the reactive α,β-unsaturated carbonyl group, copper derivatives are valued for their broad range of properties [10]. The coloration of these compounds is attributed to the chromophore (-CO-CH=CH-), whose effect depends on the presence of additional auxochromes [1]. These are polyphenolic compounds that change color and are usually yellow or orange pigments in plants due to the conjugated bonds in their structure [1,7]. Owing to the presence of two aromatic rings linked by a three-carbon α,β-unsaturated carbonyl system, these compounds can exist as either trans (E) or cis (Z) isomers [1]. Chalcones typically exist in either cis or trans configurations because of the double bond in their structure, and they generally adopt a nearly planar shape [7].

2.3. Occurrence of Chalcones in Natural Sources

In nature, chalcones are most commonly present as chalcone aglycones and chalcone glycosides, although they may also undergo modifications such as hydroxylation, condensation, or methylation [12]. Importantly, chalcones are involved in plant defense mechanisms, helping to combat reactive oxygen species and thereby ensuring plant survival while preventing molecular damage and injury caused by microorganisms, insects, and animals [8].

The most prevalent chalcones in food include glucoside, phlorizin (floretin 2’-O-glucose), naringenin (a flavonoid), and arbutin (Figure 2). Flavone and phlorizin are typically associated with apples, while naringenin is characteristic of tomatoes. Arbutin is notably present in pears, and is also found in strawberries, bearberry, wheat and wheat products, as well as in trace amounts in tea, coffee, red wine, and broccoli [13]. Prominent examples of chalcones also include butein. These compounds are widely distributed in strawberries, various berries, certain wheat products, tomatoes, pears, apples, citrus fruits, and hops (Humulus lupulus) [14].

Figure 2. Chalcones in natural products.

More specifically, they are found in a variety of fruits and vegetables, such as apples (Malus domestica) and other citrus fruits, tomatoes (Solanum lycopersicum), various plants and spices, such as licorice (Glycyrrhiza inflata), many of which have been used in traditional herbal medicine for millennia [1,15]. Flavanones, dihydroxycinnamates, and chalcones, along with their glycosides, are the main categories of polyphenolic compounds found in tomatoes [7]. Licorice is also a type of legume, rich in bioactive chalcones and belonging to the Leguminosae family. This plant is widely used in the manufacture of pharmaceutical products, such as iron pills and cough suppressants, to mask the bitter taste of other treatments [7]. In fact, such derivatives have even been found in soy-based foods [16]. Various members of the Bidens and Coreopsis genera of the Asteraceae family are also noted for their high chalcone content [17]. Their occurrence is particularly significant in the Heliantheae tribe, and several chalcones have been isolated from Helianthus annuus [18].

Chalcones are widely distributed in various parts of plants, including roots, rhizomes, leaves, and seeds [1]. They are abundant in nature, from ferns to higher plants, and many of them have polyhydroxylic acrylic rings [1]. They have also been found in the aerial roots of Ficus microcarpa, the stems and leaves of the genus Morus, the leaves or fruits of the genus Artocarpus, and members of the genus Dorstenia, all belonging to the Moraceae family [19]. In the Fabaceae family, chalcones are found in Desmodium renifolium, the roots of the Sophora genus, the Glycyrrhiza genus, and the Dalbergia genus [12]. Finally, chalcones are also found in plant species of other families, sometimes in very high concentrations, such as in the yellow juice of Angelica keiskei [20] or in members of the genus Scutellaria [12].

These edible plants are an integral part of the human diet, and the various naturally active compounds they contain, especially polyphenols, polysaccharides, and amino acids, have always been a notable topic of research among nutritionists. As precursors of polyphenolic compounds in edible plants, chalcones are not only widespread but also have diverse biological activities due to their unique structure [17].

2.4. Initial Uses/Applications of Chalcones in Agriculture and Plant Protection

With the European Union’s “From Farm to Fork” strategy aiming to reduce the use of synthetic pesticides by 2030, bioinsecticides are becoming increasingly popular not only for their different modes of action but also for their increased environmental sustainability and their applications in “copper farming” [14]. Due to their numerous biological activities, the future practical application of chalcogenides as environmentally friendly plant growth regulators and defensive agents in agriculture and plant production is noteworthy and worth discussing. In terms of pest defense and weed control, the most interesting biological properties of chalcones are their bactericidal, antifungal, anthelmintic, insecticidal, antiviral, and phytotoxic properties [12]. Modifying chalcones by adding specific functional groups can enhance their desirable properties, while at the same time, due to their unique structure, chalcones also serve as intermediates in the synthesis of therapeutically valuable compounds [14].

The chemical structures of some chalcones with potential use in agriculture and plant protection are presented in Table 1.

Table 1. Chemical structures and biological activities of some chalcones important for agriculture and plant protection.

Structure	Name	Biological Activity
	2’,4’-dihydroxy-3’-methoxychalcone	Herbicidal Action Svetaz et al. [21]
	2’,3,4,4’-tetrahydroxy-3’-geranylchalcone	Herbicidal Action Jayasinghe et al. [22]
	2’,3’,4’,4-tetrahydroxychalcone	Antimicrobial Action Onyilagha [23]
	(2E)-1-(4’-nitrophenyl)-3-(2,4,5-trimethoxyphenyl)prop-2-en-1-one	Fungicidal Action Nunes et al. [24]
	(E)-1-[2-hydroxy-4-methoxy-3-(3-methylbut-2-enyl)phenyl]-3-phenylprop-2-en-1-one	Insect Repellent Action Simmonds et al. [25]
	(E)-1-(5-hydroxy-2,2-dimethylchromen-6-yl)-3-phenylprop-2-en-1-one	Insect Repellent Action Simmonds et al. [25]
	2’,4-Dihydroxy-3’,4’-dimethoxychalcon	Phytotoxic Action Chotsaeng et al. [26]
	2’,4’,4-Trihydroxy-3’-methoxyxchalcone	Phytotoxic Action Chotsaeng et al. [26]

However, because plants produce chalcones in limited amounts, extracting them from natural sources remains difficult, and their relatively short half-life further complicates their study and application as pesticides. These challenges have driven extensive research into the synthesis and development of chalcones. There is now an urgent need for comprehensive studies to clarify their mechanisms of action, assess their effectiveness under real agricultural conditions, and evaluate their environmental and human safety [18]. Below are categories of environmentally friendly products based on chalcones.

2.4.1. Chalcones as Herbicides and Plant Growth Regulators

Exploring the chemistry of molecules naturally selected to function in ecological defense provides a promising starting point for discovering new pest control agents. Chalcones are gaining recognition in agriculture for their phytotoxic properties, which support the development of novel herbicides. Studies indicate that numerous chalcones exhibit strong herbicidal effects while showing low toxicity toward crops. As noted earlier, their activity depends on factors such as the substituents on the A and B rings of their structure (Figure 1), the concentrations applied, and the targeted plant species and organs. Derivatives with functional groups like phenoxyacetic acid, 4-(N,N-dimethylamino)phenyl, and N-methylpyrrole have shown particularly significant inhibitory effects [14].

A study by Chotsaeng [26] found that flabocavaines, xanthoxylin derivatives related to chalcones through the Claisen–Schmidt reaction as shown in Figure 3, significantly inhibited the growth of Chinese amaranth and weed [14].

Figure 3. Claisen–Schmidt condensation of xanthoxyline with aromatic aldehydes to synthesize flavokawains and chalcones, reproduced from [26].

In addition, Diaz-Tielaz in another study [27] examined the selective phytotoxic effects of chalcones on crops and weeds. Their study evaluated spray and irrigation applications on mature Arabidopsis and found that trans-chalcone adversely affected germination and early root development in certain weeds and crops, while exhibiting beneficial effects in others [14]. In additional research [28], phloretin, a well-known dihydrochalcone, demonstrated marked dose-dependent growth inhibition, pronounced morphological abnormalities, and altered behavior in Arabidopsis plants. These results further emphasize the potential of chalcones as effective and selective agents for controlling plant growth and weeds [14]. Indeed, chalcones exhibit bioactivity against nearly all eukaryotic organisms and some prokaryotic species, with numerous molecular targets. Over recent decades, intensive agriculture has relied heavily on synthetic pesticides for crop protection, often in an uncontrolled manner. This excessive use of synthetic herbicides has resulted in the emergence of agrochemical-resistant weeds, leading to significant economic losses, potential health hazards, and environmental contamination. These studies aim to identify compounds with novel mechanisms of action distinct from those of conventional synthetic agrochemicals, in order to develop effective agents with reduced environmental impact [12].

2.4.2. Chalcones as Fungicides

Numerous studies have demonstrated the efficacy of natural chalcones against a wide variety of plant pathogens responsible for serious agronomic and economic losses worldwide [12,14]. Their natural occurrence inspired the development of synthetic chalcones with enhanced antifungal properties [14], which have since become widely recognized for their activity against both plant and human fungal pathogens. Mechanistically, chalcones inhibit β(1,3)-glucan and chitin synthase, enzymes essential for the synthesis of fungal cell wall polymers, thereby compromising fungal integrity and growth [29].

Several experimental studies highlight the breadth of these effects. For example, Svetaz [21] reported that Phomopsis longicolla was highly sensitive to chalcones derived from Zuccagnia punctata. The chloroform fraction of an ethanolic extract, containing compounds such as 2’,4’-dihydroxy-3’-methoxychalcone and 2’,4’-dihydroxychalcone, exhibited strong antifungal activity against P. longicolla and Colletotrichum truncatum, two pathogens that severely affect soybean quality and yield. In a related study, Badaracco [30] demonstrated that 1,3-diphenyl-2-propen-1-one inhibited several agriculturally important fungi, including Alternaria sp., P. longicolla, Fusarium proliferatum, and Fusarium subglutinans. Likewise, Oleszek [31] showed that methanolic extracts of apple cores, particularly those enriched in phlorizin (a chalcone), were strongly active against Botrytis sp., Fusarium oxysporum, Petriella setifera, and Neosartorya fischeri [14].

Synthetic derivatives have also proven effective. Chen [32] found that pyridazine-containing chalcones displayed stronger antifungal activity than azoxystrobin, disrupting fungal cell membranes and inhibiting growth. The mechanism was associated with abnormal mycelial growth, surface rupture, and structural damage, underscoring the structural versatility of chalcones in antifungal design. Furthermore, chalcones isolated from the leaves of Myrica serrata—notably 2’,4’-dihydroxy-3’,5’-dimethyl-6’-methoxychalcone and stekurenesin—showed potent inhibition against Cladosporium cucumerinum, a common cucumber pathogen [33]. Additional chalcones from Z. punctata were also active against P. longicolla and Alternaria alternata [12]. Similarly, five chalcones isolated from Artocarpus nobilis (Moraceae) inhibited Cladosporium cladosporioides, which affects wheat, and Aspergillus niger, the causative agent of black mold in fruits and vegetables [22].

Together, these findings highlight both the natural and synthetic potential of chalcones as antifungal agents, with activities spanning multiple pathogens of agricultural importance. A summary of chalcones with reported fungicidal properties is provided in Table 2.

Table 2. Chalcones mentioned above used as insecticides are summarized in the table.

Structure	Name	Biological Activity
	Xanthohumol, 2’,4,4’-Trihydroxy-6’-methoxy-3’-(3-methylbut-2-en-1-yl)chalcone	Insecticidal Action Díaz-Tielas et al. [12]
	2’,4’-dihydroxy-3’-methoxychalcone	Fungicidal Action Molecules et al. [14]
	Isoxanthoumol 7-hydroxy-2-(4-hydroxyphenyl)-5-methoxy-8-(3-methylbut-2-enyl)-2,3-dihydrochromen-4-one	Díaz-Tielas et al. [12]
	2’,4’-dihydroxychalcone	Fungicidal Action Molecules et al. [12]
	Cordifolin, 1-(2’,3’,4’-trihydroxyphenyl)-3-(4”- methoxyphenyl)-propen-1-one]	Insecticidal Action Díaz-Tielas et al. [12]
	2’,4’-dihydroxy-3’,5’-dimethyl-6’-methoxychalcone	Fungicidal Action Oleszek et al. [31]

2.4.3. Chalcones as Insecticides

Significant research has been devoted to the synthesis and development of new chalcones and their derivatives with insecticidal properties, emphasizing structural modifications and the choice of appropriate substituents [14]. Many studies have confirmed the effectiveness of both natural and synthetic chalcones as insecticidal agents [12].

Among naturally occurring chalcones, xanthohumol and isoxanthohumol, isolated from hops (Humulus lupulus L.), serve as notable examples. These compounds have shown considerable insecticidal activity against the peach–potato aphid (Myzus persicae) [34].

The research of Shakil and Saxena [35] reported the isolation of a novel chalcone, cordifoline, from the woody stem of Tinospora cordifolia and assessed its effects on Spodoptera litura larvae. Their results indicated that cordifoline delayed larval development, extended the larval period, and reduced larval weight [14]. Meanwhile, Hidalgo [36] investigated both bis- and mono-chalcones against Spodoptera frugiperda, finding that two mono-chalcones containing brominated and hydroxyl groups on the A ring and an N,N-dimethyl group on the B ring caused larval mortality rates of 40% and 60%, respectively [14].

The research conducted by Kumar et al. [37] is especially notable as it represents the first documented case of chalcones synthesized via microwave irradiation exhibiting pesticidal activity against Plutella xylostella [14]. Their study revealed that an electron-withdrawing group on the A ring is essential for pesticidal effectiveness, while the B ring can accommodate either electron-withdrawing or electron-donating substituents. Chlorine (-Cl) substituents and their positions within both rings were found to be particularly important. Among the compounds tested, 1,3-bis(4-chlorophenyl)prop-2-en-1-one exhibited the highest pesticidal activity, laying the groundwork for further structural optimization and the development of chalcone-based pesticides targeting P. xylostella and related insect pests [14].

Moreover, the common hop, Humulus lupulus L. (Cannabaceae), contains more than 1000 distinct chemical compounds [38]. The flavonoids present in hops (H. lupulus L.) are the focus of extensive research due to their beneficial health effects [34]. Xanthohumol, the principal chalcone in hop cones at an approximate concentration of 1%, possesses numerous valuable biological activities [39].

Growing concerns about the health risks associated with the extensive use of pesticides and synthetic fertilizers have driven increased interest in safer and more eco-friendly plant protection solutions. As a result, agricultural practices increasingly favor the use of naturally occurring pesticidal compounds from plants as alternatives to harmful chemical pesticides [40]. These compounds not only eliminate harmful microflora and pests but are also safe for both humans and the environment, which is a primary concern. Being natural substances, they are completely biodegradable. Furthermore, they exhibit activity against specific insect groups. This makes them environmentally friendly agents that can influence insect taste receptors, deterring feeding on plants and ultimately causing starvation and death [34].

3. Recent Advances on the Observed Health Promoting Properties of Chalcones

Over the last ten years, there has been a significant increase in interest in copper nitrates due to their interesting biological activities. As mentioned above, chalcones have a fantastic compound that allows them to produce a wide variety of new heterocyclic compounds with interesting pharmacological effects. In fact, they have shown promising therapeutic efficacy in treating a variety of disorders due to a wide range of structural modifications. This is because, in fact, scientific studies have confirmed that chalcone derivatives, based on their structure, have a special connection with such a wide range of pharmacological actions [14]. Structural modifications of copper rings have led to a high degree of diversity that has proven useful for the development of these new pharmaceutical agents, making them a subject of ongoing interest in both academia and industry [6]. Over recent decades, extensive research has been carried out on the pharmacological properties of both natural and synthetic chalcones, including their anti-inflammatory, antioxidant, anti-infective (such as anti-leukemic, anti-malarial, and anti-tuberculosis), antiviral, and particularly anticancer activities. Indeed, several chalcones have been successfully developed into commercial drugs for treating certain digestive system disorders, while others are currently undergoing clinical trials for applications in cancer therapy, cardiovascular disease management, and the treatment of viral infections [12,41]. Some of the main useful properties of chalcones are listed below.

3.1. Antioxidant Properties of Chalcones

As noted, the high presence of chalcones in foods such as fruits, vegetables, and medicinal plants underscores their potential role as natural antioxidants. Antioxidants are compounds that slow down or prevent oxidation, a process that generates free radicals. These free radicals can trigger chain reactions that damage cells, contributing to oxidative stress. Such stress is linked to the development of chronic conditions, including heart disease, strokes, cancer, arthritis, respiratory disorders, Parkinson’s disease, and various inflammatory ailments [9]. Chalcones demonstrate notable antioxidant potential owing to their chemical composition and effectiveness, with their electron-rich phenolic structure making them particularly well-suited as antioxidant agents. Among their various properties, antioxidant activity is arguably the most straightforward feature of chalcones [42].

Chalcones have been shown to influence the activity of antioxidant enzymes and regulate gene expression, thereby strengthening their capacity to protect against oxidative damage. Their ability to interact with multiple pathways linked to oxidative stress positions them as promising candidates for therapeutic approaches in health maintenance and disease prevention. Oxidative stress inflicts significant damage on nucleic acids, lipids, proteins, and vital enzymes, contributing to the development of conditions such as cancer, arthritis, cardiovascular disorders, and neurodegenerative diseases [42].

Antioxidant chalcones act through multiple cellular targets and mechanisms. They boost enzymatic antioxidant defenses, including superoxide dismutase, catalase, and glutathione peroxidase, and activate the Nrf2-ARE pathway to enhance the expression of antioxidant and detoxification genes. Additionally, chalcones can suppress the generation of reactive oxygen species (ROS) by inhibiting enzymes such as NADPH oxidases and xanthine oxidases, and they possess metal-chelating abilities that help prevent ROS formation catalyzed by metals [3,4].

There are many chalcones with different antioxidant properties that even follow different metabolic pathways [43].

3.1.1. Butein

3,4,2’,4’-tetrahydroxycoumarin, also known as butein, is a natural coumarin isolated mainly from the tree Toxicodendron vernicifluum, which is traditional and widely known in China [43]. The ability of butein to positively modulate the expression of nuclear factor erythroid 2-related factor 2 (Nrf2), a critical transcription factor involved in the regulation of many cellular pathways [44]. Primarily, Nrf2 directly influences key regulators such as C/EBP-α and PPARγ, which, as mentioned earlier, contribute to preserving the differentiated state of preadipocytes [45]. Conversely, Nrf2 serves as a vital transcription factor essential for sustaining the redox balance in tissues [44].

Specifically, Nrf2 controls the expression of numerous enzymes linked to oxidative stress, including antioxidant enzymes such as thioredoxin, glutathione, heme oxygenase-1 (HO-1), and quinone oxidoreductase (NQO1), as well as detoxifying enzymes like glutathione S-transferases (GSTs) [46]. Overall, HO-1 is recognized as a crucial enzyme in antioxidant processes. It catalyzes the rate-limiting step in the breakdown of free heme into iron, carbon monoxide, and biliverdin, all of which are metabolites possessing significant antioxidant properties [47]. Recently, several studies have highlighted the critical role of HO-1 in the management of metabolic homeostasis, given its high level of expression in the white adipose tissue of genetically obese mice induced by a high-fat diet (HFD) [48]. Consequently, Nrf2 and its downstream enzyme HO-1 could serve as potential therapeutic targets for obesity-related disorders. In this context, Yang et al. demonstrated that treating 3T3-L1 cells with butein results in elevated Nrf2 expression, which subsequently modulates HO-1 mRNA expression levels [44]. A similar experimental design was followed by Wang et al., who also confirmed the ability of butein to induce increased expression of HO-1 mRNA and related protein expression in the 3T3-L1 adipocyte cell line [49]. Furthermore, Song et al. carried out a comparative investigation into the anti-obesity potential of several natural compounds derived from Rhus verniciflua Stokes, a lacquer tree long valued in traditional medicine.

3.1.2. Panduratin A

Panduratin A (PAN A) is a flavonoid featuring a chalcone backbone with three oxygenated groups located solely on the B ring and a geranyl substituent at the C2–C3 position formed via the Diels–Alder reaction. This natural chalcone is primarily obtained from Boesenbergia pandurata, a traditional medicinal plant renowned for its antioxidant and anti-inflammatory effects. In this regard, a study explored the potential of PAN A as a novel AMPK activator [4]. Modulating AMPK represents a promising strategy for addressing obesity, given its critical role as an energy sensor in mammalian cells. In their study, the authors observed that treating various cell models (including 3T3-L1 adipocytes, HepG2 liver carcinoma cells, and L6 skeletal muscle cells) with PAN A led to AMPK-dependent inhibition of ACC, thereby reducing endogenous lipid synthesis. Specifically, AMPK activation can be triggered by several regulators such as LKB1, CaMKKβ, sirtuin (SIRT1), and NAD(P)H. To elucidate the molecular mechanism behind PAN A’s activation of AMPK, the researchers examined its effects on the expression of these activators. They discovered that PAN A treatment enhanced the translocation of LKB1 from the nucleus to the cytoplasm and increased its binding affinity to AMPK, resulting in rapid enzyme activation [4,45].

3.1.3. 4’-OH-Flurbiprofen-Chalcone

Alzheimer’s disease (AD) is the most common type of dementia, which is a fatal, chronic, neurodegenerative disease of the brain [48]. However, because Alzheimer’s disease (AD) involves complex pathological mechanisms, current therapeutic approaches—such as cholinesterase inhibitors and N-methyl-D-aspartate (NMDA) receptor antagonists—primarily address symptoms and are unable to effectively halt or reverse the progression of neurodegeneration [46].

Although the exact pathogenesis of Alzheimer’s disease (AD) remains unclear, several factors—including β-amyloid (Aβ) accumulation, oxidative stress, and reduced acetylcholine levels—have been identified as key contributors to its development. A sustained imbalance between the production and clearance of Aβ1-42 results in the buildup of Aβ1-42 monomers, oligomers, and eventually large insoluble amyloid fibrils [47]. Amyloid deposits can therefore lead to cerebrovascular damage, neuroinflammation, abnormal calcium homeostasis, and neurodegeneration. Therefore, reducing the accumulation of Ab1-42 is a potential therapeutic strategy for treating AD [50].

A recent study demonstrated that chalcones possess properties suitable for Aβ imaging tracers, exhibiting high brain uptake and strong affinity for Aβ aggregates. Moreover, multifunctional agents containing phenolic hydroxyl groups showed enhanced antioxidant activity [48]. Consequently, 4’-OH-flurbiprofen was chosen for combination with chalcones to create a series of 4’-OH-flurbiprofen–chalcone hybrids, designed to function as multifunctional agents with both inhibitory effects on Aβ aggregation and antioxidant properties [50].

The in vitro antioxidant activities of the 4’-OH-flurbiprofen–chalcone hybrids were evaluated using the ORAC-FL method (oxygen radical absorbance capacity with fluorescein). Trolox (6-hydroxy-2,5,7,8-tetramethylchromane-2-carboxylic acid), a water-soluble analogue of vitamin E, served as the standard, and antioxidant activity was expressed in Trolox equivalents. Among the tested compounds, 4’-hydroxy-flurbiprofen and 4-(diethylamino)chalcone exhibited the strongest antioxidant activities. Notably, 4-(diethylamino)chalcone also demonstrated significant radical-scavenging potential. All hybrid compounds showed higher ORAC-FL values compared to flurbiprofen alone, indicating that the introduction of the chalcone unit and the free 4’-OH group into flurbiprofen was crucial for enhanced antioxidant activity. Hybrids containing two phenolic hydroxyl groups displayed greater antioxidant capacity than those with only one. Furthermore, hybrids with dimethylamino or diethylamino groups at position 4’ of the chalcone moiety exhibited remarkable antioxidant properties. Thus, both the phenolic hydroxyl groups and the amino substituents at position 4’ were essential for improving antioxidant activity [50].

Chalcones with antioxidant properties are summarized in Table 3 below.

Table 3. Chalcones with antioxidant properties.

Structure	Name	Biological Activity
	3,4,2’,4’-tetrahydroxychalcone, Butaine	Antioxidant Action, Annie-Mathew et al. [43]
	Panduratin A	Antioxidant Action Maisto [4]
	4-(Diethylamino)chalcone	Antioxidant Action, N. Guzior et al. [50]
	4-(Dimethylamino)chalcone	Antioxidant Action, N. Guzior et al. [50]

3.2. Health Promoting Properties of Chalcones Against Atherosclerosis and CVDs

Accumulating preclinical data indicate that natural and synthetic chalcones exert cardioprotective effects across complementary mechanisms relevant to atherothrombosis. In vascular and macrophage models, representative chalcones (e.g., xanthohumol, licochalcone A, cardamonin, panduratin A) reduce NF-κB-driven inflammation, attenuate ROS generation via Nrf2/HO-1 activation, and modulate lipid handling (↓oxLDL uptake; ↑cholesterol efflux) [51]. In animal models of atherosclerosis and cardiometabolic risk, chalcones have been associated with improvements in endothelial function (↑eNOS/NO bioavailability), reductions in vascular oxidative stress and cytokines (TNF-α, IL-6), and decreases in lesion burden or lipid indices, alongside antiplatelet/antithrombotic actions (inhibition of platelet activation/aggregation and thrombus formation) [5]. Additional benefits have been reported in ischemia–reperfusion and cardiac remodeling settings, where chalcones can limit infarct size, fibrosis, and hypertrophy via modulation of PI3K/Akt, MAPK, and apoptotic signaling [52]. Early human data remain limited; small exploratory studies and nutraceutical formulations suggest acceptable short-term tolerability and signal-level effects on inflammatory/oxidative markers, but robust randomized trials evaluating hard cardiovascular outcomes are still lacking [53]. Overall, the weight of evidence supports chalcones as promising multi-target leads for anti-inflammatory, antioxidant, endothelial-protective, and antithrombotic strategies in atherosclerosis and CVD, while underscoring the need for standardized pharmacokinetic optimization, dose-ranging, and controlled clinical evaluation [52].

3.2.1. Anti-Atherosclerotic and Cardioprotective Effects of Chalcones

In recent years, coronary heart disease has accounted for around 7.3 million deaths, while high blood pressure and stroke have caused approximately 9.4 million and 6.2 million deaths, respectively. Obesity and atherosclerosis continue to be the primary underlying pathological factors contributing to these conditions [49]. Projections indicate that by 2030, cardiovascular-related deaths could rise to approximately 23.3 million. The majority of these diseases are attributed to modifiable risk factors, including smoking, poor dietary habits, lack of physical activity, hypertension, obesity, diabetes, and elevated lipid levels [54].

The growing prevalence of fast food consumption, unhealthy dietary habits, and sedentary lifestyles has made cardiac disorders—especially atherosclerosis and associated progressive diseases—a major concern within the medical community [49]. Lower-income, middle-income countries are disproportionately affected by cardiovascular disease (~75%), and the number of deaths is almost equal in men and women. The reason for the decline in cardiovascular disease in developed countries may be improved healthcare facilities and better access to newer drugs [54]. The global demand for novel and more effective cardiovascular agents remains a top priority in managing heart disease, as cardioprotection has been a central focus of research over the past 10–15 years. However, an ideal cardio-protective compound or drug has yet to be clearly identified [49].

Several natural and semi-synthetic chalcones have emerged as promising candidates for the inhibition of various cardiovascular disorders. Chalcone-based compounds that exhibit strong biological activity and possess well-defined mechanisms of action (MOAs) and structure–activity relationships (SARs) can serve as valuable prototypes for developing antihypertensive, anti-angiogenic, antiarrhythmic, and cardioprotective drugs. By leveraging insights into these molecular targets, structural characteristics, and SARs, researchers can focus on designing chalcone derivatives that are more potent, selective, safe, and cost-effective for cardiovascular therapy [49].

Over 4000 polyphenolic compounds have existed in the plant kingdom for more than a billion years. These compounds are widely present in fruits, vegetables, tea, and wine, and are generally classified into nine subgroups, including flavonols, flavones, flavanones, flavanols, isoflavones, anthocyanidins, and proanthocyanidins, many of which exhibit promising cardioprotective properties [55]. Several chalcones, such as tinctorimine, lonchocarpin, xanthohumol, xanthohumol B, desmethylxanthohumol, xanthoangelol, xanthoangelol E, isobachalcone, derricin, hydroxysaflor yellow A, 4-hydroxyderricin, hydroxylated chalcones, substituted chalcone imides, sulfonamide-substituted chalcones, and lupulone-based chalcones, have been identified as bioactive agents targeting cardiovascular systems [49].

A list of chalcones, their cardiovascular targets, and their physicochemical properties is presented in Table 4. The cardioprotective effects of chalcones are closely linked to their structural features. The α,β-unsaturated carbonyl group is essential for bioactivity, as it enables Michael addition reactions with nucleophilic sites on target proteins involved in oxidative and inflammatory pathways. Substitutions on the aromatic rings strongly influence potency: hydroxyl groups, particularly in ortho and para positions, enhance antioxidant and radical-scavenging properties, while methoxy groups can increase lipophilicity and membrane permeability. Prenylated chalcones, such as xanthohumol and its derivatives, show improved bioavailability and stronger inhibition of platelet aggregation and vascular inflammation. Halogenated chalcones have demonstrated superior activity in ischemia–reperfusion models, likely due to increased stability and interactions with cardiomyocyte signaling proteins. In contrast, glycosylation often reduces activity, reflecting diminished cell permeability. Collectively, these SAR insights highlight how relatively small modifications of the chalcone scaffold can markedly alter their antioxidant, anti-inflammatory, and antithrombotic activities, offering guidance for the rational design of novel chalcone-based cardiovascular agents.

It has recently been found that a few natural compounds (such as curcumin (bis-chalcone), quercetin (flavonoid), provide adequate protection against myocardial infarction (MI) and other cardiovascular diseases such as hypertension, hyperlipidemia, thromboembolism, and arrhythmia through their anti-dyslipidemic and antioxidant action, the attenuation/inhibition of lipid peroxidation, increased expression of cardioprotective proteins, and through various other pathways [56]. Among these compounds, a few chalcones and their hybrids are reported to exert a protective effect against myocardial infarction caused by ischemia/reperfusion. These chalcones significantly reduce the size of the infarction, moderate lipid peroxidation, and reduce/inhibit protein expression [49].

A study evaluated the cardioprotective potential of halogenated chalcones (Cl- and F-substituted chalcones) against myocardial infarction caused by ischemia/reperfusion (I/R) in rats using 2,3,5-triphenyl tetrazolium chloride as a staining agent. The study revealed that chalcones significantly reduced infarct size and lipid peroxidation, supporting their cardioprotective activity against MI [56]. Similar cardioprotective effects were observed with YLSC. The authors reported that YLSC reduced infarct size, serum LDH, and AST. It also reduced the apoptosis rate and protein expression of GRP78 and caspase 12 in the myocardium [57].

In addition to existing and established chalcones, a few 1,3-diphenyl-2-E-propen-1-one derivatives have been found to exhibit remarkable activity in inhibiting various cardiovascular targets. Chalcones such as 3,2’,4’,6’-tetrahydroxy-4,3’-dimethoxychalcone, 2-hydroxychalcone, and neovavachalcone have shown usefulness in reperfusion of ischemic injuries, response to vascular damage, regression of vascular damage, and prevention of coronary atherosclerosis. 2’,4’-dihydroxy-6’-methoxychalcone, lycopene B, and isoliquiritigenin are promising candidates for the management of hypertension and the prevention of MI and stroke through their β-adrenergic receptor blocking action. Similarly, mixed receptor blocking action (alpha- and beta-adrenergic receptors) has been demonstrated by glypalichalcone and licochalcone A. Interestingly, antithrombotic properties have been reported for licochalcone G, which effectively inhibits coagulation factor Xa. Licochalcone A and G, glypalichalcone, neobavachalkone, 3,2’,4’,6’-tetrahydroxy-4,3’-dimethoxychalcone, and 2’,4’-dihydroxy-6’-methoxychalcone are molecules with impressive anti-obesity activity. Table 4 summarizes some chalcones with anti-atherosclerotic activity [57].

Table 4. List of various copper compounds with cardiovascular action.

Structure	Name	Biological Activity
	3,2’,4’,6’-tetrahydroxy-4,3’-dimethoxy chalcone	Anti-cardiovascular diseases Sivasankaran et al. [58]
	2-hydroxychalcone	Anti-cardiovascular diseases Lecour, K. T. Lamont [59]
	Neobavachalcone	Anti-cardiovascular diseases Annapurna et al. [60]
	2’,4’-dihydroxy-6’-methoxychalcone	Anti-cardiovascular diseases Jian et al. [57]
	Isoliquiritigenin	Anti-cardiovascular diseases Zhou et al. [61]
	Curcumin	Anti-cardiovascular diseases Mahapatra, K. Bharti [49]
	Tinctormine	Anti-cardiovascular diseases Mahapatra, K. Bharti [49]
	Lonchocarpin	Anti-cardiovascular diseases Mahapatra, K. Bharti [49]
	Derricin	Anti-cardiovascular diseases Mahapatra και S. K. Bharti [49]

Table 4. List of various copper compounds with cardiovascular action.

Structure	Name	Biological Activity
	3,2’,4’,6’-tetrahydroxy-4,3’-dimethoxy chalcone	Anti-cardiovascular diseases Sivasankaran et al. [58]
	2-hydroxychalcone	Anti-cardiovascular diseases Lecour, K. T. Lamont [59]
	Neobavachalcone	Anti-cardiovascular diseases Annapurna et al. [60]
	2’,4’-dihydroxy-6’-methoxychalcone	Anti-cardiovascular diseases Jian et al. [57]
	Isoliquiritigenin	Anti-cardiovascular diseases Zhou et al. [61]
	Curcumin	Anti-cardiovascular diseases Mahapatra, K. Bharti [49]
	Tinctormine	Anti-cardiovascular diseases Mahapatra, K. Bharti [49]
	Lonchocarpin	Anti-cardiovascular diseases Mahapatra, K. Bharti [49]
	Derricin	Anti-cardiovascular diseases Mahapatra και S. K. Bharti [49]

3.2.2. Health Benefits of Chalcones Against Heart Failure

Almost all cardiovascular diseases lead to changes in the structure and function of the heart [58]. The systolic and diastolic functions of the myocardium are essential for maintaining the heart’s pumping ability, with calcium serving as a critical ion in initiating electromechanical coupling in the heart [59]. In end-stage heart failure, environmental disturbances disrupt the stability of intracellular ion channels, leading to calcium accumulation that can trigger malignant arrhythmias. Mitochondria, as the main source of cellular energy, are particularly affected by calcium overload, which induces oxidative stress and impairs mitochondrial functions, thereby compromising cardiomyocyte performance [61]. Flavonoids help regulate the expression of Cav1.2 and NCX1 plasma membrane transporters, maintaining calcium homeostasis in cardiomyocytes and protecting against mitochondrial apoptosis. Through reducing oxidative dysfunction and preventing mitochondrial permeability transition, flavonoids can counteract the progression of heart failure [61]. Myocardial fibrosis is a hallmark of advanced heart failure, characterized by collagen fiber deposition and interstitial cell proliferation that disrupts the heart’s structure and accelerates pump failure. Transforming growth factor beta (TGF-β) plays a significant role in fibrosis pathogenesis, with TGF-β1 closely associated with collagen production in the heart. Flavonoids exhibit therapeutic effects akin to antioxidants, offering anti-fibrotic benefits by regulating key factors such as Nrf2 and NF-ĸB. For instance, quercetin modulates the expression of the antioxidant enzyme Prx-3 by regulating Nrf2 transcription factors, thereby reducing oxidative damage to mitochondria [55,56].

In addition, the monomer 17-methoxy-7-hydroxy-benzoin-furachalcone plays an important role in alleviating heart failure by targeting key mechanisms involved in cardiac remodeling and dysfunction. This chalcone promotes the expression of endothelial nitric oxide synthase (eNOS) and nitric oxide (NO) by activating the PI3K/Akt pathway, which is vital for maintaining vascular health and reducing oxidative stress. By enhancing NO production, chalcone helps improve endothelial function, reduce inflammation, and prevent structural remodeling of the heart [62]. Cardiac remodeling, a hallmark of advanced heart failure, often involves fibrosis, oxidative stress, and reduced myocardial function [63]. 17-Methoxyl-7-hydroxy-benzene-furanchalcone counteracts these effects by maintaining mitochondrial function, inhibiting oxidative damage, and maintaining calcium homeostasis within cardiomyocytes. These actions collectively mitigate mitochondrial dysfunction, calcium overload, and inflammation, which contribute significantly to the progression of heart failure [62].

There is sufficient evidence for the positive effects of flavonoids on cardiac function in patients with heart failure, as well as their ability to reverse drug-related heart damage, supporting the therapeutic potential of flavonoids for clinical heart failure.

3.3. Chalcones in Clinical Trials Against CVDs and Other Inflammation-Related Disorders

Over recent decades, chalcones have attracted significant attention as bioactive compounds due to their diverse pharmacological properties, which include anti-inflammatory, anticancer, antimicrobial, antiviral, and dermatological effects. Structurally characterized by an α,β-unsaturated carbonyl system, chalcones act as key precursors in the biosynthesis of flavonoids and are naturally present in a wide range of edible plants, herbs, and traditional medicinal sources. Given their structural versatility and biological relevance, increasing efforts have been made to investigate chalcones not only in vitro and in vivo but also in human clinical settings.

The following Table 5 presents a detailed summary of clinical and ex vivo studies conducted on different chalcone derivatives. It highlights the purpose of each study, the type and number of participants or test organisms, the experimental design, and the key outcomes observed. Compounds such as xanthohumol, cardamonin, panduratin A, flavokawain A, and others have been evaluated for their effects on inflammation, cancer progression, viral infections, skin health, metabolic disorders, and more. These findings contribute significantly to understanding the translational potential of chalcones from bench to bedside. The diversity of methods—ranging from double-blind placebo-controlled trials to targeted molecular pathway analysis—illustrates the growing scientific interest in integrating chalcone-based compounds into therapeutic and nutraceutical applications.

Table 5. Clinical Trials with Chalcones against CVDs and other inflammation-related disorders.

Name of Chalcone	Aim of the Study	Type of Organism Used/Number and Type of Volunteers	Study Design	Main Findings	Reference/Year of Study
Xanthohumol	Determine if xanthohumol has anti-inflammatory effects in healthy humans if applied in low doses achievable through dietary intake.	Human 14 healthy young men and women	Quantity (of Chalcone) Dose administered: 0.125 mg xanthohumol per volunteer control group: Received a placebo beverage Time (Sampling & Stimulation Timing) Blood sample timing: Before and 1 h after beverage intake PBMC stimulation duration: 24 h and 48 h post-isolation LTA stimulation: Applied to PBMCs post-isolation and to HEK293 cells in vitro Method (Molecules & Assays Used) Cells used: Peripheral Blood Mononuclear Cells (PBMCs) HEK293 cells transfected with human TLR2 (hTLR2) Stimulus applied: Lipoteichoic acid (LTA)	Oral ingestion of low-dose xanthohumol (0.125 mg) altered LTA-induced immune responses in monocytes from healthy individuals. Xanthohumol suppressed CD14/TLR2-dependent activation of immune cells. The findings suggest xanthohumol can reduce inflammatory signaling triggered by LTA.	[64] (July 2020)
Panduratin A	To evaluate the clinical effects of Boesenbergia pandurata intake on skin hydration, gloss, wrinkle formation, and elasticity.	Humans The trial included 92 human subjects receive either B. pandurata ethanol extract (BPE) or a placebo for 12 weeks.	A double-blind, placebo-controlled trial was conducted to clinically assess the effects of Boesenbergia pandurata ethanol extract (BPE), containing 8% panduratin A, on human skin hydration, gloss, wrinkle formation, and elasticity.	The test group showed a significant increase in skin hydration and gloss after 12 weeks compared to the placebo group. The test group showed a significant reduction in skin wrinkling compared to the placebo group. Although there was no statistically significant difference in skin elasticity between the groups, the test group exhibited a greater improvement rate. The results suggest that B. pandurata ethanol extract (BPE) may be effective as a nutraceutical or nutricosmetic for improving skin hydration, gloss, and reducing wrinkles.	[65] (April 2017)
Cardamonin	To investigate the ex vivo immunomodulatory effects of cardamonin on pro-inflammatory cytokine production in the context of primary Sjögren’s syndrome (pSS).	Peripheral blood mononuclear cells (PBMCs) Cells from patients diagnosed with pSS and from healthy controls	Cells were exposed to different concentrations of cardamonin to assess its anti-inflammatory effects. Inflammatory Markers: TNF-α and IL-6 levels were measured by ELISA, while nitric oxide (NO) production was evaluated using the Griess method. Mechanism of Action: iNOS expression and NF-κB activity were analyzed via immunofluorescence staining to understand the underlying regulatory pathways.	Cardamonin effectively inhibited the production of TNF-α, IL-6, and NO, and also downregulated iNOS expression and NF-κB activation. Cardamonin has a significant immunomodulatory effect on inflammatory pathways in pSS, highlighting its potential benefit as a therapeutic candidate for controlling inflammation associated with the disease.	[66] (January 2018)
Flavokawain A	Explore the role of PRMTs (Protein arginine methyltransferases) regulators of protein function via arginine methylation and their potential as therapeutic targets in cancer. Determine the mechanism of action of Flavokawain A (FKA) in bladder cancer (BC), focusing on its interaction with PRMTs. Assess whether FKA could serve as a candidate PRMT inhibitor for future bladder cancer therapy	Human bladder cancer cells Mouse xenograft models	Examined PRMT5 expression in surgically removed bladder cancer (BC) specimens to understand its role in cancer progression. Used bioinformatics tools (computational simulation, virtual screening, molecular docking, energy analysis) and co-IP/mutation assays to investigate how Flavokawain A (FKA) interacts with PRMT5. Applied bio-layer interferometry, CETSA, and pull-down assays to confirm the direct binding between FKA and PRMT5. Conducted cell-based assays and mouse xenograft models to assess the anti-cancer effects of FKA and validate its potential as a PRMT5-targeting therapeutic in bladder cancer.	FKA, a natural product, is a novel small-molecule inhibitor of PRMT5. It was shown to inhibit breast cancer (BC) progression both in vitro and in vivo by targeting PRMT5. FKA exhibited strong anti-BC activity, supporting its potential for further development toward clinical application.	[67] (October 2022)
4’ ethoxy-2’-hydroxy-4, 6’ dimethoxy- chalcone (Ro 09-0410)	To evaluate the antirhinovirus activity of this “pro-drug” in volunteers, a double-blind, placebo-controlled study was conducted in which oral Ro 09-0415 was administered both before and after exposure to a virulent strain of human rhinovirus.	Human Healthy volunteers	Double-blind placebo-controlled trial volunteers were given Ro 09-0415 or placebo orally before and after nasal challenge with rhinovirus type 9. Clinical symptoms, virus isolation, antibody response, and drug levels were measured over the course of the trial	Plasma concentrations of the active compound exceeded those needed to inhibit rhinovirus type 9 in vitro. No evidence was found to suggest that treatment with Ro 09-0415 provided a beneficial effect. It is unlikely that oral Ro 09-0415 will be effective in treating human rhinovirus infection.	[68] (1984)
licochalcone A in combination with 4-t-butylcyclohexanol	to develop an active ingredient concept for the treatment of sensitive skin tested compounds regarding their potential to (i) decrease the release of proinflammatory mediators, (ii) counteract the hyperresponsiveness of nerve fibres and, thus, exert effects on cutaneous neurosensory dysfunction.	In vitro (on cells): The study tested compounds on human skin cells (fibroblasts, TRPV1-expressing cells) and pig neurons to evaluate anti-inflammatory and nerve-calming effects. In vivo (on humans): Two controlled studies were done on human volunteers to test creams for reducing skin stinging and shaving-induced redness.	In vivo tests (on humans): Two controlled, single-blind, randomized studies were conducted. Tested 4-t-butylcyclohexanol, alone or combined with licochalcone A, for relieving skin sensitivity symptoms (e.g., stinging, redness). In vitro tests: Evaluated 4-t-butylcyclohexanol, licochalcone A, and acetyl dipeptide-1 cetyl ester. Assessed their ability to: Reduce PGE2 release and NFκB activation (inflammatory markers). Inhibit TRPV1 activation and CGRP release (nerve-related responses).	In vitro findings: Licochalcone A effectively reduced NFκB signaling and PGE2 secretion at lower concentrations than acetyl dipeptide-1 cetyl ester. In vivo findings (on humans): A formulation combining 4-t-butylcyclohexanol and licochalcone A-rich licorice extract significantly reduced shaving-induced redness (erythema).	[69] (February 2016)
Xanthohumol	To evaluate whether a micellar formulation of xanthohumol improves its oral bioavailability compared to native xanthohumol in humans.”	Humans Healthy males and females	double-blind, crossover trial a single dose of 43 mg was orally administered as a native or micellar formulation.	Micellar xanthohumol significantly improved absorption compared to native xanthohumol: 5× higher total exposure (AUC) of its main metabolite (xanthohumol-7-O-glucuronid). Over 20× higher peak plasma concentration of the metabolite. Unmetabolized (free) xanthohumol in the blood remained very low (~1% or less), indicating rapid metabolism after absorption.	[70] (September 2018)
Hydroxysafflor Yellow A	To study how safe Hydroxysafflor Yellow A (HSYA) is, how well it is tolerated, and how it behaves in the human body (pharmacokinetics).	Humans 12 healthy chinese volunteers	Part A was a randomized, double-blind, placebo-controlled dose-escalation study that evaluated safety and tolerability Part B was an open-label study where 12 volunteers received 75 mg of HSYA once daily for 14 consecutive days to examine PKs. Form: Pure HSYA powder for injection. Administration: Intravenous infusion (into the vein). Dosage schedules: Single doses ranging from 6.25 mg to 125 mg Multiple doses: 100 mg and 125 mg once daily for 7 days Part B: 75 mg once daily for 14 days	HSYA was well tolerated across all doses PK analysis revealed rapid absorption, no significant accumulation, and effective elimination.	[71] (January 2025)
Licochalcone A, l-carnitine and 1,2-decanediol	To compare the tolerability and efficacy of moisturizers containing licochalcone A, L-carnitine, and 1,2-decanediol (active formulation) with a placebo in treating mild to moderately severe acne in Asian subjects.	Humans 120 Asian patients	An 8-week double-blind, prospective, randomized controlled study was conducted. All patients were equally randomized into three groups: (A) adapalene gel, (B) adapalene gel combined with the active formulation, and (C) adapalene gel combined with a placebo. Acne severity, skin bioengineering parameters, and skin tolerability were assessed throughout the study.	The active formulation group showed significant reductions in inflammatory lesions. Skin irritations were observed less frequently than in the other two groups. The combined use of adapalene gel and the moisturizer reduced undesirable side effects. The moisturizer may be superior to placebo in preventing cutaneous irritations and improving patients’ adherence to acne treatment.	[72] (September 2015)
6-prenylnaringenin (6-PN) 8-prenylnaringenin (8-PN)	To compare the oral bioavailability and safety of 6-PN and 8-PN and investigated their effects on peripheral blood mononuclear cells (PBMCs).	Humans 16 Healthy young women and men,	double-blind, placebo-controlled, crossover trial a single oral dose of 500 mg 6-PN, 8-PN, or placebo in random order.	8-PN is significantly more bioavailable in healthy humans than its isomer 6-PN. Despite its lower bioavailability, 6-PN is similarly effective as 8-PN in enhancing PBMC viability.	[73] (March 2018)
Xanthohumol	To investigate the anticancer effects of xanthohumol with a focus on breast cancer. aimed to clarify the effectiveness of XN in preventing or reducing breast cancer growth and to understand the molecular mechanisms involved, particularly focusing on the Notch signaling pathway and apoptosis regulation.	4T1 breast tumor mouse model two human breast cancer cell lines—MCF-7 and MDA-MB-231	In a 4T1 breast tumor mouse model, xanthohumol (XN) treatment was evaluated for its impact on tumor growth using tumorigenicity assays and immunohistochemistry to assess markers like Notch1 and Ki-67. In vitro, MCF-7 and MDA-MB-231 human breast cancer cells were treated with xanthohumol (XN); effects on cell viability, apoptosis, and cell cycle were assessed using MTT assay, flow cytometry, and Western blot to analyze apoptosis-related and Notch signaling proteins.	Xanthohumol (XN) significantly inhibited tumor growth in a breast cancer mouse model and reduced cell viability, induced G0/G1 cell cycle arrest, and promoted apoptosis in cultured breast cancer cells. These effects were confirmed by flow cytometry and Western blot, showing decreased Bcl-2/Bcl-xL expression and altered caspase-3 activity, indicating apoptosis. The Notch signaling pathway was notably inhibited by XN in both in vivo and in vitro models, highlighting its role in suppressing tumor progression.	(November 2017) [74]

4. Conclusions—Future Perspectives

Chalcones constitute a structurally diverse group of flavonoids with well-documented antithrombotic, anti-inflammatory, and antioxidant properties that contribute to cardiovascular protection. By modulating key molecular pathways such as NF-κB, Nrf2/HO-1, and PI3K/Akt, chalcones can suppress vascular inflammation, inhibit platelet aggregation, and enhance endothelial function—mechanisms that are central to the prevention and management of atherosclerosis and cardiovascular diseases.

Structure–activity relationship (SAR) studies have shown that subtle chemical modifications, including hydroxylation, methoxylation, and prenylation, significantly influence biological activity and pharmacokinetic behavior. These findings underscore the potential of chalcones as multi-target molecular scaffolds for the rational design of new cardioprotective and anti-inflammatory therapeutics.

Despite encouraging preclinical results, clinical evidence remains limited. Future research should therefore prioritize: (i) well-designed clinical trials to validate efficacy and safety in humans; (ii) improvement of bioavailability and stability through nanocarrier and formulation technologies; (iii) structure-guided optimization to enhance potency, selectivity, and pharmacokinetic profiles; and (iv) expanded investigation of chalcones in sclerosis-related and other inflammation-driven disorders, where their antifibrotic and antioxidant actions may offer additional benefits.

In summary, chalcones represent a promising class of bioactive compounds with significant potential in the development of next-generation therapeutic and nutraceutical agents targeting atherosclerosis, cardiovascular disease, and other chronic inflammatory conditions. Advancing this field will require integrated interdisciplinary efforts that bridge synthetic chemistry, molecular pharmacology, and clinical research to fully translate their pharmacological promise into clinical reality.