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REVIEW article

Front. Pharmacol., 02 March 2021 | https://doi.org/10.3389/fphar.2020.578970

Indian Medicinal Plants and Formulations and Their Potential Against COVID-19–Preclinical and Clinical Research

  • 1Bioactive Natural Product Laboratory, Department of Pharmacognosy and Phytochemistry, School of Pharmaceutical Education and Research, Jamia Hamdard (Deemed University), New Delhi, India
  • 2Centre for Translational and Clinical Research, Jamia Hamdard (Deemed University), New Delhi, India
  • 3Department of Biosciences, Jamia Millia Islamia (Central University), New Delhi, India
  • 4Department of Surgery, School of Unani Medical Education and Research, Jamia Hamdard (Deemed University), New Delhi, India

The cases of COVID-19 are still increasing day-by-day worldwide, even after a year of its first occurrence in Wuhan city of China. The spreading of SARS-CoV-2 infection is very fast and different from other SARS-CoV infections possibly due to structural differences in S proteins. The patients with severe diseases may die due to acute respiratory distress syndrome (ARDS) caused by systemic inflammatory reactions due to the excessive release of pro-inflammatory cytokines and chemokines by the immune effector cells. In India too, it is spreading very rapidly, although the case fatality rate is below 1.50% (https://www.statista.com), which is markedly less than in other countries, despite the dense population and minimal health infrastructure in rural areas. This may be due to the routine use of many immunomodulator medicinal plants and traditional AYUSH formulations by the Indian people. This communication reviews the AYUSH recommended formulations and their ingredients, routinely used medicinal plants and formulations by Indian population as well as other promising Indian medicinal plants, which can be tested against COVID-19. Special emphasis is placed on Indian medicinal plants reported for antiviral, immunomodulatory and anti-allergic/anti-inflammatory activities and they are categorized for prioritization in research on the basis of earlier reports. The traditional AYUSH medicines currently under clinical trials against COVID-19 are also discussed as well as furtherance of pre-clinical and clinical testing of the potential traditional medicines against COVID-19 and SARS-CoV-2. The results of the clinical studies on AYUSH drugs will guide the policymakers from the AYUSH systems of medicines to maneuver their policies for public health, provide information to the global scientific community and could form a platform for collaborative studies at national and global levels. It is thereby suggested that promising AYUSH formulations and Indian medicinal plants must be investigated on a priority basis to solve the current crisis.

1 Introduction

A novel coronavirus-induced pneumonia, which was later called coronavirus disease 2019 (COVID-19), has rapidly increased to an epidemic scale and affected whole human population globally (WHO, 2020a). In India, the first case of COVID-19 was an imported case from Wuhan, China on January 30, 2020 traced in Kerala (Sahasranaman and Kumar, 2020) and the death rate of COVID-19 in India is 1.45%, as of 12th December, 2020 (Worldometers, 2020). Severe acute respiratory syndrome-related coronavirus (SARS-CoV-2) has become a pandemic hazard to global public health worldwide.

Coronaviruses (CoVs) are large viruses comprising of four genera, namely alpha, beta, gamma, and delta. The beta-coronavirus class includes severe acute respiratory syndrome (SARS) virus (SARS-CoV), Middle East respiratory syndrome (MERS) virus (MERS-CoV), and the COVID-19 causative agent SARS-CoV-2. (Li G. et al., 2020). The novel SARS-CoV-2 is a beta CoV that shows 88% similarity to two bat-derived SARS-like CoVs (bat-SL-CoVZC45 and bat-SL-CoVZXC21), about 50% identical to the sequence of MERSCoV, and 70% similarity in genetic sequence to SARS-CoV (Cheng and Shan, 2020). Although there is an extremely high resemblance between SARS-CoV and the novel SARS-CoV-2, the SARS-CoV-2 is spreading rapidly as compared to the SARS-CoV, which may be explained by structural differences in the S proteins (Rabaan et al., 2020).

The SARS-CoV-2 S protein has been found as a significant determinant of virus entry into host cells using angiotensin converting enzyme 2 (ACE2) receptor similar to SARS-CoV. Whereas the binding affinity of virion S glycoprotein and ACE2 is reported to be 10–20 folds higher in SARS-CoV-2 as compared to that of SARS-CoV (Song et al., 2018).

Severe cases of COVID-19 are reported to have increased plasma concentrations of pro-inflammatory cytokines, including interleukins (IL-6 and IL-10), tumor necrosis factor (TNF)-α granulocyte-colony stimulating factor (G-CSF), monocyte chemoattractant protein 1 (MCP1), and macrophage inflammatory protein (MIP)1α (Yuki et al., 2020). Akin to the common viral infections, the antibody profile against the SARS-CoV virus manifests a typical pattern of IgM and IgG antibody production. The IgG antibody is believed to play a protective role, as the SARS-specific IgG antibodies last for a longer time while IgM antibodies disappear at the end of 12 weeks. The latest reports show a significant reduction in the number of CD4+ and CD8+ T cells in the peripheral blood of SARS-CoV-2-infected patients, besides activation of other pro-inflammatory cytokines such as nuclear factor-κB (NF-κB), interferon regulatory factor 3 (IRF3) and type I Interferons (IFN-α/β) (Li G. et al., 2020). A recent report shows that many patients died from acute respiratory distress syndrome (ARDS) caused by the cytokine storm, which is a deadly uncontrolled systemic inflammatory response resulting from the release of large amounts of pro-inflammatory cytokines and chemokines by immune effector cells in SARS-CoV infection (Guo et al., 2020).

Although the pathogenesis of COVID-19 is still not clear, patients with COVID-19 show non-specific symptoms ranging from no symptoms (asymptomatic) to severe pneumonia and death. However, the most common symptoms include fever, non-productive cough, dyspnea, myalgia, fatigue, diarrhea, lung damage, normal or decreased leukocyte counts, and radiographic evidence of pneumonia, which are similar to the symptoms of SARS-CoV and MERS-CoV infections (WHO, 2020b; Rothan and Byrareddy, 2020). Complications include ARDS, acute heart injury, and secondary infections (Guo et al., 2020).

The present conventional strategy of the disease control includes isolation of cases and tracing their contacts, providing optimal care to these infected cases, reducing chances of secondary infections by early diagnosis, and rapid development of effective diagnostic, preventive and therapeutic strategies, including vaccines (WHO, 2020b). The treatment approach for COVID-19 is supportive care, which is supplemented by the combination of broad-spectrum antibiotics, antivirals, corticosteroids and convalescent plasma (Yang et al., 2020).

Scientists are working hard to develop effective treatments. As of October 18, 2020, more than 3611 clinical trials (with more than 100 complementary medicines) on COVID-19 are either ongoing or enrolling patients, and new ones are being added every day, as the case count skyrockets globally. The drugs being tested range from repurposed flu treatments to failed ebola drugs, to malaria treatments that were first developed decades ago (Lythgoe and Middleton, 2020). There is scale-up development of vaccines across the world by many pharmaceutical companies as well as research organizations. These treatments undergoing trials may require months or years to develop and hit the market, meaning that an immediate treatment or control mechanism should be found, if possible (Table 1).

TABLE 1
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TABLE 1. Details of clinical trials completed on AYUSH drugs for COVID-19 (Source: www.ctri.nic.in).

Considering the current situation, various treatment modalities have been well-thought-out, including traditional medicine, which has been widely used during the past epidemic outbreaks, including SARS and H1N1 influenza (Luo et al., 2020). Until now three countries including India, China, and South Korea, have issued guidelines on traditional regimens for the prevention and management of COVID-19 (Ang et al., 2020).

The Indian Traditional System of Medicine is one of the oldest systems of medical practice in the world and has played an essential role in providing health care service to human civilization, right from its inception. India has the exclusive distinction of its own recognized traditional medicine; Ayurveda, Yoga, Unani, Siddha, and Homoeopathy (AYUSH) (Adhikari and Paul, 2018). These systems are based on definite medical philosophies and represent a way of achieving a healthy lifestyle with conventional and established ideas on the prevention of diseases and the promotion of health. The basic treatment approach of all these systems is holistic and the pharmacological modalities are based on natural products of plants, animals, or mineral origin. Given this, there is a resurgence of interest in AYUSH systems, which have helped the nation in the pandemic crisis due to plague, cholera, Spanish flu, etc. in the past. Hence, by repurposing the traditional uses of Indian medicinal plants and formulations, new treatment options can be identified to combat the current deadly pandemic. In view of the COVID-19 outbreak, the entire human race across the globe is perturbed. While there is no medicine for COVID-19 as of now, it is imperative to take preventive measures such as practicing self-hygiene, social distancing and boosting immunity. Many safe traditional formulations of AYUSH, which are well known immunity modulators, have been used for centuries in respiratory disorders and in allergic conditions. The Ministry of AYUSH (Govt of India) has listed out such formulations and recommended their use as a prophylactic measure in red zones, containment zones, as well as for corona warriors. Many of them are now under clinical trial in COVID-19 patients (Table 1).

TABLE 2
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TABLE 2. AYUSH recommended prophylactic approach through Ayurvedic formulations. Ref: AYUSH Ministry of Health Corona Advisory-D.O. No. S. 16030/18/2019- NAM; dated: 06th March, 2020. Ref: AYUSH Ministry of Health Corona Advisory -F.No. Z 25.23/09/2018–2020-DCC (AYUSH); dated: 24th April, 2020.

TABLE 3
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TABLE 3. AYUSH recommended prophylactic approach through Unani formulation. Ref: AYUSH Ministry of Health Corona Advisory–D.O. No. S. 16030/18/2019- NAM; dated: 06th March, 2020.

TABLE 4
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TABLE 4. AYUSH recommended prophylactic approach through formulations of Siddha system of medicine. Ref: AYUSH Ministry of Health Corona Advisory–D.O. No. S. 16030/18/2019-NAM; dated: 06th March, 2020.

Similarly, there are many medicinal plants indigenous to India and used in the Indian Systems of Medicine which have been reported as potent antiviral with immunomodulatory and anti-allergic/anti asthmatic activities. Many of these medicinal plants are also an integral part of several traditional formulations that have been in use for a long time.

This review discusses the possible alternative strategies for the management of the SARS-CoV-2 infection by reducing its morbidity in patients as an adjuvant to modern therapy and also by providing prophylactic management. Further, potential testing targets of botanicals from Indian medicinal plants need to be explored against SARS-CoV-2 infection and categorized on a priority basis in view of their reported antiviral, immunomodulatory and other related activities.

2 Potential Traditional Indian/AYUSH Formulations for the Management of COVID-19

There is plenty of data supporting the effectiveness of herbs in treating the viral infection. For instance, in controlling the contagious disease spread in the Guangdong Province of China during the 2003 SARS outbreak (Zhang et al., 2020). There are convincing pieces of evidence to establish that traditional Chinese medicine (TCM) has favorable effect in the treatment or prevention of SARS (Yang et al., 2020). A combination of modern and traditional therapy might reduce the severity of the disease, intensity of symptoms, death rate, and side effects. Similar are the observations for Shuanghuanglian (A Chinese medicine) a liquid composed of a blend of honeysuckle, Chinese skullcap, and forsythia, which is claimed to have antiviral, antibacterial, and immunomodulatory effects (https://www.bioworld.com/). Since AYUSH encompasses five different systems of medicine, rich in a variety of traditional formulations, it is likely to have a better chance than other systems to come up with a satisfactory solution to the COVID-19 crisis.

Ayurveda means ‘Science of life’. It provides a complete system to have a long and healthy life. It is derived from the concepts of “Dinacharya” - daily regimes and “Ritucharya” - seasonal regimes to maintain a healthy life. Uplifting and maintaining the immunity is duly emphasized across the Ayurveda’s classical scriptures.

The Unani system of medicine, known as Greco-Arab Medicine, is built on the four conditions of living (hot, sodden, frosty, and dry) and four humors of Hippocratic hypothesis namely, blood, yellow bile, dark bile, and mucus. Epidemics, referred to as waba in the Unani system of medicine, are thought to occur if any contagion or ajsam-i -khabitha, finds a place in air and water. Furthering the view, Ibn-e-Sina (980–1035 CE) stated that epidemics spread from one person to another, and one city to another ‘like a message’ (Sina, 1878).

AYUSH systems of medicine propagate general preventive measures aimed at preventing the spread of infection such as social distancing, hygiene and anti-septic measures (sanitization of surroundings), improvement of immunity, and promotion of general health (dietary modifications and herbal drugs). The present article elucidates some traditional Indian AYUSH formulations with proven antiviral, anti-asthmatic, and immunomodulatory activities, however their role in combating COVID-19 needs to be established. Clinical trials of AYUSH medicines like Ashwagandha, Yashtimadhu, Guduchi, Pippali, and AYUSH-64 on patients, health workers, and those working in high-risk areas have been initiated in India by the Ministry of AYUSH, Ministry of Health and Family Welfares, and the Council of Scientific and Industrial Research (CSIR) with the technical support of Indian Council of Medical Research (ICMR) (Table 1).

3 AYUSH Recommendations for Management of COVID-19

Based on the different systems of Indian Medicine, separate recommendations have been issued from time to time from the Ministry of AYUSH (Government of India) for the management of COVID-19. These different approaches are being followed by the Hospitals as per their specialization, mainly as adjuvants to modern medicine, which could be potentially relevant for COVID 19 treatment. Details of recommended formulations are described below and depicted in Table 2 (Ayurveda), Table 3 (Unani) and Table 4 (Siddha).

3.1 Ayurvedic Approaches

3.1.1 AYUSH Kwath

Ministry of AYUSH promotes the use of AYUSH kwath, which is a ready-made formulation for health promotion of the masses. The formulation is made of four herbs Ocimum sanctum L. leaves, Cinnamomum verum J. Presl. stem barks, Zingiber officinale Roscoe rhizomes and Piper nigrum L. fruits. The formulation is sold in the market with different names like ‘AYUSH Kwath’, ‘AYUSH Kudineer’ or ‘AYUSH Joshanda’. It is available in powder and tablet forms in the market. These herbs are reported to boost immunity (Carrasco et al., 2009; Niphade et al., 2009; Alsuhaibani and Khan, 2017; Bhalla et al., 2017) and are active remedies to various viral diseases (Mair et al., 2016; Ghoke et al., 2018; Pruthvish and Gopinatha, 2018).

3.1.2 Samshamani Vati

Samshamani vati (Guduchi ghana vati) is an ayurvedic formulation used in all types of fevers. It is also used as an antipyretic and anti-inflammatory remedy (Patgiri et al., 2014). Samshamani vati is made of aqueous extract of Tinospora cordifolia (Willd.) Miers (family Menispermaceae), and reported to be an immunomodulator (More and Pai, 2011) due to the synergistic effect of the various compounds present. It is also effective in various viral diseases (Sachan et al., 2019).

3.1.3 AYUSH-64

AYUSH-64 tablet is composed of Alstonia scholaris (L.) R. Br. bark, Picrorhiza kurroa Royle ex Benth. rhizomes, Swertia chirayita (Roxb.) H. Karst. whole plant, and Caesalpinia crista L. seed pulp. Because of its antimalarial activity, AYUSH-64 is considered to be effective among the high-risk coronavirus population. Researchers have reported that each of its constituents is effectively antiviral, anti-asthmatic, and immunoboosting (Sharma et al., 1994; Siddiqui et al., 2012; Sehgal et al., 2013; Panda et al., 2017; Win et al., 2019; Woo et al., 2019).

3.1.4 Agasthaya Hareetaki

Agastya Haritaki Rasayana is a popular ‘Avaleha kalpana’, used in the management of various respiratory infection and comprises more than 15 herbal ingredients. Most of its ingredients showed antiviral, anti-asthmatic, anti-inflammatory, and immunomodulatory activities (Mouhajir et al., 2001; Tripathi and Upadhyay, 2001; Balasubramanian et al., 2007; Vadnere et al., 2009; Patel and Asdaq, 2010; Pathak et al., 2010; Jain et al., 2011; Kumar et al., 2011; Lampariello et al., 2012; Jiang et al., 2013). The above literature suggests the symptomatic management of COVID-19 by Agastya Haritaki.

3.1.5 Anuthaila

Anuthaila consists of about twenty ingredients and out of them Leptadenia reticulate (Retz.) Wight and Arn. has been reported in allergic response, treatment of asthma, bronchitis, and throat trouble (Mohanty et al., 2017). Similarly, Ocimum sanctum L. is recommended for a wide range of conditions including, cough, asthma, fever, and malaria (Cohen, 2014) and Sesamum indicum L. oil for dry cough, asthma, migraine, and respiratory infections (Nagpurkar and Patil, 2017). There are reports on S. indicum seeds with Tachyspermum ammi (L.) Sprague seeds for dry cough, asthma, lung diseases, and common cold (Patil et al., 2008). On the basis of above literature, Anuthaila justifies its use in corona virus pandemic condition (Table 2).

3.2 Unani Approaches

3.2.1 Triyaq-e-Araba

Triyaq-e-Araba is an important Unani formulation used as a detoxifying agent. It contains Laurus nobilis L. berries, Bergenia ciliate (Haw.) Sternb. stem, Aristolochia indica L. roots and Commiphora myrrha (Nees) Engl. It has been reported by several authors as a potent antiviral agent (Aurori et al., 2016), including against SARS-CoV (Loizzo et al., 2008). Further, B. ciliata is found to be effective against the influenza virus-A and herpes simplex virus-1 (HSV-1) (Rajbhandari et al., 2003), whereas its active principal, bergenin, has been found to be effective against hepatitis C virus (HCV) and HIV virus (Ahmad et al., 2018). On the basis of this literature, Triyaq-e-Araba could be one of the effective antiviral medicine and certifies its use against COVID-19.

3.2.2 Roghan-e-Baboona

Roghan-e-Baboona is an Unani remedy utilized as an anti-asthmatic and for the treatment of inflammatory complaints. Flowers of Matricaria chamomilla L. are the main ingredient of Roghan-e-Baboona. It is composed of the flowers of M. chamomilla, which is found effective for acute viral nasopharyngitis (Srivastava et al., 2010), as well as for sore throat (Kyokong et al., 2002).

3.2.3 Arq-e-Ajeeb

Arq-e-Ajeeb is a liquid preparation that contains thymol, menthol, and camphor. Thymol is a promising candidate for topical application as an antiviral agent for herpetic infections (Lai et al., 2012; Sharifi-Rad et al., 2017). Menthol has been reported as an anti-inflammatory agent (Zaia et al., 2016). The Unani physicians have a very successful history of treating Nazla wabai (Swine flu) using Arq -e-Ajeeb. These studies support the use of Arq-e-Ajeeb for COVID-19.

3.2.4 Khamira-e-Banafsha

Khamira-e-Banafsha is a semi-solid Unani formulation prepared by adding decoction of flowers of Viola odorata L. to a base of sugar or sugar with honey and used for cold-cough as expectorant and for the treatment of ailments of respiratory system and chest diseases, bronchitis, whooping cough, fever, expectorant, antipyretic etc. Further, V. odorata has been reported to suppress the viral load and increase antiretroviral drug efficacy (Gerlach et al., 2019), decrease the thickness of the alveolar wall, hemorrhage area, and alter the epithelial lining of bronchioles of the lungs (Koochek et al., 2003). The above literature supports its use for the management of COVID-19.

3.2.5 Laooq-e-Sapistan

Laooq-e-Sapistan is a semisolid sugar-based polyherbal Unani formulation extensively used by the masses in India for the treatment of cold and cough, whooping cough, and phlegm. It reduces inflammation of the pharynx, tonsils, and irritation or infection. The jelly like sticky mass of ripe fruit of Cordia myxa L. is the main ingredient, which has been reported as antiviral and antitussive (Jamkhande et al., 2013). Another important constituent is Ziziphus fruit, which contains betulinic acid. Literature showed the down-regulation of IFN-γ level by betulinic acid in mouse lung, thus enhancing immunity and suggested as potential therapeutic agent for viral infections (Hong et al., 2015). Aqueous extract also reported increasing thymus and spleen indices as well as enhance the T-lymphocyte proliferation, hemolytic activity, and natural killer (NK) cell activity (Yu et al., 2016). Viola odorata L., one of its ingredients, suppresses the viral load (Gerlach et al., 2019). Hence, the literature supports the use of AYUSH formulation Laooq-e-Sapistan in COVID-19.

3.2.6 Sharbat-e-Sadar

Sharbat-e- Sadar is an Unani polyherbal syrup formulation and is widely used for common cold, cough and respiratory diseases. Trachyspermum ammi (L.) Sprague, an important ingredient, reported to neutralize antibodies for Japanese encephalitis virus (Roy et al., 2015), and a glycoprotein was found to proliferate B-cells (Shruthi et al., 2017). Adhatoda vasica Nees inhibits HIV-Protease (Singh et al., 2010), Bombyx mori was reported to increase immune responses against viral infection (Lü et al., 2018). Other ingredients such as Glycyrrhiza glabra L., Ficus carica L., Onosma bracteatum Wall., and Ziziphus jujuba Mill. also possess the antiviral and immunomodulatory activities, as summarized in Table 5.

TABLE 5
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TABLE 5. List of Indian Medicinal Plants/AYUSH drugs with proven immunomodulatory, antiviral and anti-allergic/anti-inflammatory/anti-asthmatic activity having potential for exploring against COVID 19 categorized for prioritization on the basis of their earlier reports.

3.2.7 Khameera Marwareed

Khameera marwareed is a compound, sugar-based, semisolid Unani formulation used as an immunomodulator. It has been reported to stimulate the immune system through T helper 1 (Th1) type cytokine response and maintains the body in a healthier position to fight against viral infections (Khan et al., 2009). Its ingredients showed powerful antiviral activities by inhibiting replication (Benencia and Courrèges, 1999).

3.2.8 Asgandh Safoof

Asgand (Withania somnifera (L.) Dunal) is a very popular Indian medicinal plant. The root powder is used in the Unani system of medicine as an immunomodulator. It is reported that the root’s extract significantly increases the CD4+ and CD8+ counts (Bani et al., 2006) and blood profile, especially WBC and platelet counts (Agarwal et al., 1999). Aqueous suspension showed potent inhibitory activity toward mitogen-induced proliferative response of T-lymphocytes and prevent SARS-CoV-2 entry by disturbing connections between viral S-protein receptor binding domain and host ACE2 receptor (Balkrishna et al., 2020). The above literature supports the preventive use of Asgandh safoof against COVID-19.

3.2.9 Habb-e-Bukhar

Habb-e-Bukhar is a polyherbal tablet formulation of Unani system of medicine, prescribed in elephantiasis and malarial fever. The main ingredient of Habb-e-Bukhar is cinchona bark. Its active constituent quinine is being used by some countries as either experimental treatment or suggested as a drug with a promising profile against COVID-19 (Devaux et al., 2020). Another constituent, Tinospora cordifolia (Willd.) Miers is reported as potent antiviral agent against HSV (Pruthvish and Gopinatha, 2018) as well as suggested for immune-enhancing activity (Rastogi et al., 2020). Thus, literature supports Habb-e-Bukhar in the treatment of COVID-19.

3.2.10 Sharbat-e-Toot Siyah

Sharbat-e-Toot Siyah is composed of the juice of Morus nigra L. in a sugar base and is used to treat tonsillitis and sore throat. It has been reported as anti-inflammatory and analgesic and inhibits the pro-inflammatory cytokines (Chen et al., 2016). Very recently, it has been reported to enhance immunomodulatory activity (Lim and Choi, 2019).

3.2.11 Laook-e-Katan

Laook-e-Katan is a sugar-based semisolid Unani formulation composed of Linum usitatissimum L. seed, which contains alpha linolenic acid and has been reported to have antiviral, anti-inflammatory, and immunomodulatory activities (Leu et al., 2004; Erdinest et al., 2012; Miccadei et al., 2016). In Unani, it is recommended for respiratory disorders (Table 3).

3.3 Siddha Approaches

3.3.1 Nilavembu Kudineer

Nilavembu Kudineer is a polyherbal Siddha formulation prescribed for the prevention and management of viral infections and fevers. It acts as an immunomodulator and plays a defending role against dengue fever and chikungunya. Recent studies showed that formulation has antiviral and antimicrobial actions, which makes it suitable for viral fevers, malaria, and typhoid fever (Mahadevan and Palraj, 2016). Previously, studies proved that most of its constituents are effective as antiviral, anti-asthmatic, and immunobooster agents (Carrasco et al., 2009; Wang et al., 2010; Jin et al., 2011; Chang et al., 2013; Wintachai et al., 2015; Mair et al., 2016).

3.3.2 Ahatodai Manapagu

Ahatodai Manapagu is composed of Adathoda vasica Nees leaves, which contains alkaloids like vasicine, the active ingredient in various cough syrups. A. vasica has been used in the Indian medicinal system for thousands of years, to treat various types of respiratory disorders (Sampath Kumar et al., 2010). Vinothapooshan et al. suggested that its extract positively modulates the immunity of the host (Vinothapooshan and Sundar, 2011).

3.3.3 Kabasura Kudineer

Kabasura Kudineer is a traditional formulation used in the Siddha system of medicine for managing common respiratory complaints such as flu and cold. Siddha practitioners also recommended this formulation for severe phlegm, dry cough, and fever. It is made up of more than ten herbal ingredients, and each ingredient has a unique pharmacological activity in respiratory disorders. Hence, the ministry of AYUSH recommends its use for symptomatic management in COVID-19 (Sampath Kumar et al., 2010; Jin et al., 2011; Vinothapooshan and Sundar, 2011; Chang et al., 2013).

In addition to Ayurvedic, Unani, and Siddha formulations recommended by AYUSH there are some homeopathic formulations such as Arsenium album, Brayonia alba, and Rhus toxicodendrum have been recommended which have not been included due to controversies over the use of homeopathic medicine. These formulations are prepared by dilutions in such a way so that no single detectable molecule is present in the final formulation, which results in controversy (Ernst, 2010). The criticism is due to nonevidential rationale to determine the biological effects of solutions containing unmeasurable starting material (Kaur, 2013).

Further, advancements in pathogenesis and understanding of diseases provide a wider platform to report the pharmacological limitations and opportunities of these highly diluted homeopathic medicines. Day by day, it is becoming more challenging for a pharmacologist to validate the therapeutic claims of homeopathic medicines through experiments. Low acceptance of homeopathic formulations is due to the absence of standardized protocols to justify their pharmacological potential. A major concern is to develop evidence-based validated methods and advancements in the homeopathic system to justify its measurable dilutions, which will help in understanding the mechanism of action and acceptability of homeopathic medicine (Table 4).

3.4 Routinely Used Common Indian Medicinal Plants for Exploring Against COVID-19

Ashwagandha, giloe, ginger, cinnamon, tulsi, black pepper, black cumin, amla, turmeric, garlic, and flax seeds have been traditionally used as herbal remedies for multiple diseases since ancient times. These herbs have been utilized in food preparations and traditional medicines in several countries. However, in India, their culinary use is very common and they are a part of kitchen in every house. Similarly, there are some traditional Indian formulations such as Chyawanprash, Triphala, and Rooh Afza etc. that are very commonly used in Indian territory as a part of daily used nutritional supplements. These plants and formulations are very common and at least one of them is being used daily by every Indian, irrespective of religion/community/financial status. The above-mentioned herbs and formulations have been proved potent scientifically for their immunomodulatory, antioxidant, and anti-infective properties, which might be one of the reasons behind the lower death rate of Indians per million of population due to COVID-19 even with minimum health infrastructure.

3.4.1 Allium sativum L. (Garlic)

Various research has been conducted in vivo to highlight the effect of A. sativum in immunomodulation using garlic oil extract. The results showed reduction in serum TNF-α, ICAM-1 and immunoglobulin (G and M) levels confirming the enhancement in immune system activity (Kamel and El-Shinnawy, 2015). Pre-treatment with aqueous garlic extract showed notable antiviral effects mainly by reduction in infectivity and titer of virus against the velogenic strain of Newcastle disease virus in embryonated chicken eggs (Arify et al., 2018). A. sativum also showed antiviral effect against avian influenza virus H9N2 on Vero cells (Rasool et al., 2017). Its defensive effect on allergen-induced airway inflammation in rodent model showed significant reduction in inflammatory cell count, eosinophil infiltration and serum IgE modulation of Th1, Th2, and Th3 cytokines, upregulation of Th-1, Th-3 and simultaneous down-regulation of Th-2 expression. (Hsieh et al., 2019). Old extract of A. sativum showed modulation of airway inflammation established in BALB/c mice by reduction in percentage of eosinophil, lavage and serum IgG1 levels, and perivascular inflammation. The study suggested the attenuation of allergic airway inflammation by aged garlic extract (Zare et al., 2008). It has been found that fresh raw garlic extract showed anti-inflammatory effects by decreasing production of prostaglandin E2 (PGE2), IL-6, IL-1β, nitric oxide (NO), and leukotrienes (LT D4 and E4) in lipopolysaccharide activated RAW264.7 cells (Jeong et al., 2016).

3.4.2 Cinnamomum verum J.Presl. (Cinnamon) or Cinnamomum zeylanicum Blume

C. verum essential oil and powder exhibited anti-oxidant, immunostimulant, and antiviral activity in Newcastle disease virus in chickens mainly by modulating total protein, globulin, total antioxidant capacity, and lysozyme activity, and significantly increased phagocytic activity (Islam et al., 2017). Another study reported that C. zeylanicum essential oil when blended with other essential oils showed effective antiviral potential against H1N1 and HSV1 viruses. Reduction in virus infectivity has been observed with 99% at 60-min contact time and more than 99.99% after 60 min for both H1N1 and HSV1 viruses (Brochot et al., 2017). Its bark extract exhibited immunomodulatory activity and significantly increased serum immunoglobulins, phagocytic index, neutrophil adhesion, and antibody titer (Niphade et al., 2009). Procyanidine polyphenols (Type A) extracted from C. zeylanicum bark showed anti-inflammatory potential in edema induced by carrageenan (Vetal et al., 2013). Alcoholic extract of bark suppressed intracellular release of TNF-α (murine neutrophils) and leukocytes (pleural fluid) as well as inhibition of TNF-α gene expression in lipopolysaccharide-stimulated human peripheral blood mononuclear cells (Joshi et al., 2010).

3.4.3 Curcuma longa L. (Turmeric)

Aqueous extract of C. longa decreased relative spleen weight and modulation in hematological changes indicating the potential of C. longa as an immunomodulator in cyclophosphamide-immunosuppressed in vivo model. The study observed promising effects of turmeric as an immunomodulator by representing spleen cells in younger mice (Mustafa and Blumenthal, 2017). C. longa extract also showed antiviral potential against dengue virus in in vitro and in vivo studies on Huh7it-1 cells and a remarkable reduction in viral load has been observed by in in vivo model (Ichsyani et al., 2017). Water and ethanolic crude extracts have been found to be antiviral in H5N1 also showed upregulated TNF-α as well as IFN-β mRNA expression, highlighting its promising role in the inhibition of the replication of viruses (Sornpet et al., 2017). Turmeric extract has been found to be anti-allergic in mice immunized with ovalbumin and alum. Attenuation of food allergy by maintaining balance of Th1/Th2 has been reported. Extract has been found to cause reduction in Th2 and increase in Th1 cell-related cytokines. Further, increased levels of IgE, IgG1 and mMCP-1 levels were also decreased proving effects of turmeric in allergic disorders mainly, asthma and food allergies (Shin et al., 2015). Various other studies also reported anti-inflammatory effects of C. longa either alone or in combination (Lee et al., 2020).

3.4.4 Linum usitatissimum L. (Flax Seed)

Heteropolysaccharide, extracted from flax seed hull possessed immunomodulatory activity and anti-hepatitis B virus potential. It significantly stimulated mRNA expression of TNF-α, NO and IL exhibiting immune responses in murine macrophages. Antiviral activity has been reported through inhibition of expression of surface antigen as well as envelop antigen and also interfered with DNA replication. The study suggested its promising potential as an immunostimulant and vaccine adjuvant (Liang et al., 2019). It showed anti-inflammatory and immunomodulatory potential in obesity-associated insulin resistance. Its oil in co-culture with 3T3-L1 adipocytes-RAW 264.7 macrophages of C57BL/6 mice reported shifting the cytokines toward anti-inflammatory with a decrement in TNF-α. Immunomodulation has been observed through an increase in levels of Th2-related cytokine (IL-4), serum anti-ova IgG1, and IgE, and a decrease in Th-1 related cytokines (TNF-α and IFN-γ) and anti-ova IgG levels (Palla et al., 2015). Another study reported the immunomodulatory activity of phenolic components of flax seed mainly through reduction in cell-mediated immune responses (Kasote et al., 2012).

3.4.5 Nigella sativa L. (Black Cumin)

Nigella sativa L.’s bioactive compounds have been observed as potential inhibitors of COVID-19 in molecular docking studies. Nigellidine gave energy complex at active site (6LU7) with energy scores closest to chloroquine and better than hydroxychloroquine and favipiravir whereas α-hederin gave energy complex at the active site (2GTB) with energy scores better than chloroquine, hydroxychloroquine, and favipiravir (Salim and Noureddine, 2020). The alcoholic seed extract has shown immunosuppressive activity on a phytohemagglutinin and immunostimulating effect on non-phytohemagglutinin (PHA) stimulated proliferation (Alshatwi, 2014). The thymoquinone-rich oil showed suppression of cytokine signaling molecules, and PGE2 in T-lymphocytes as well as enhanced PGE2 release in adrenocarcinomic human alveolar basal epithelial A549 cells (Koshak et al., 2018).

3.4.6 Ocimum sanctum L. (Tulsi)

Hydro-alcoholic extract of Ocimum sanctum inhibited intracellular multiplication of virus. It also inhibits non-specific interference with virus-cell interactions in H9N2 viruses. (Ghoke et al., 2018). The immunomodulatory potential of alcoholic leaves extracts at IC50 value of 73.3 μg/ml showed reduction in hepatic parasite and, skewing of the humoral response toward Th1 type (Bhalla et al., 2017). O. sanctum inhibits leukotriene-C4-synthase, leukotriene-A4-hydrolase and cyclooxygenase-2 activities in cultured HL-60 cells and causes a significant reduction in OVA-induced lung inflammation (Soni et al., 2015).

3.4.7 Phyllanthus emblica L. (Amla)

Amla has been reported to significantly relieve chromium-induced immunosuppressive effect on lymphocyte proliferation and led to restoration in production of IL-2 and INFγ (Sai Ram et al., 2002). Phenolics from emblica has been found to increase splenocytes proliferation. Geraniin and isocorilagin showed significant immunostimulatory effects (Liu et al., 2012). Ethanolic extract of amla strongly reduced levels of pro-inflammatory cytokines and increased levels of anti-inflammatory cytokine (Bandyopadhyay et al., 2011). An isolated compound (1, 2, 4, 6-tetra-O-galloyl-β-d-glucose) of P. emblica showed antiviral potential against HSV by HSV-1 inactivation, which leads to inhibition of early infection indulging attachment and penetration of virus, suppression of intracellular growth and inhibited gene expression of HSV-1 E and L along with DNA replication (Xiang et al., 2011).

3.4.8 Piper nigrum L. (Black Pepper)

Piperamides isolated from P. nigrum fruits showed significant inhibition of coxsackie virus type B3 in a cytopathic effect inhibition assay (Mair et al., 2016). Aqueous extract of P. nigrum acted as a potent modulator of the macrophages and significantly enhanced splenocyte proliferation in a dose-dependent manner (Majdalawieh and Carr, 2010). The isolated alkaloid from P. nigrum exhibited anti-inflammatory effect in RAW 264.7 cells stimulated by LPS and significant inhibition in iNOS-mediated NO and IL-1β, IL-6, and TNF-α. It also demonstrated anti-inflammatory activity in edema induced by carrageenan (Pei et al., 2020). Reports have confirmed the improvement of ovalbumin-induced nasal epithelial barrier dysfunction in allergic rhinitis mouse model. Further, protection of epithelium integrity, enhancement in E-cadherin tight junction protein as well as inhibition of the degraded levels of zonula occludens-1 and occluding in the nasal passage have been reported. Additionally, enhancing the activation of Nrf2/HO-1 signaling showed anti-allergic and anti-asthma activities (Bui et al., 2020).

3.4.9 Tinospora cordifolia (Willd.) Miers (Giloe)

In vitro screening of T. cordifolia silver nanoparticles against chikungunya virus cell showed significant antiviral potential (Sharma V. et al., 2019). Alcoholic leaves extract of T. cordifolia significantly decreases intracellular reactive oxygen species (ROS) in chikungunya patients with high levels of intracellular ROS in persisting polyarthralgia by ex vivo treatment (Banerjee et al., 2018). An in vitro study revealed the antiviral potential of crude stem extract of T. cordifolia against HSV in Vero cell lines by inhibiting the growth of HSV (Pruthvish and Gopinatha, 2018). Aqueous extract of T. cordifolia stem significantly increase INFγ and IL levels (IL-1, IL-2, IL-4) in isolated chicken peripheral blood mononuclear cells (PBMCs) against infectious bursal disease virus. Further, immunomodulatory potential via the toll like receptor (TLR)-mediated pathway was also concluded (Sachan et al., 2019). The hydro-alcoholic extract of T. cordifolia stem in drinking water caused enhancement of cellular immunity as well as humoral immunity in broiler chicks (Nety et al., 2017). Chloroform extract significantly prevented pro-inflammatory biomarkers (IL-6, IL-1β and PGE2) and decreased paw oedema (p ≤ 0.05) with no toxicity reported when conducted in RAW264.7 macrophages (Philip et al., 2018).

3.4.10 Withania somnifera (L.) Dunal (Ashwagandha)

Multiple studies have proved that Ashwagandha has antiviral and immunomodulatory potential. Very recently, an in silico study concluded that Withaferin-A exhibits antiviral potential against SARS-CoV-2 through inhibiting RNA polymerase with higher binding energy than hydroxychloroquine and other drugs used against SARS-CoV-2. Another study on withanone showed blockage of SARS-CoV–2 entry and also its subsequent infection by interrupting electrostatic interactions between the RBD and ACE2 (Balkrishna et al., 2020). Grover and colleagues through molecular docking reported the potential of withaferin A against HSV through inhibition of DNA polymerase enzyme (Grover et al., 2011). W. somnifera molecular mechanism has been elucidated by using network ethnopharmacological technique and reported that withanolide-phytosterol combination is a good immunomodulator (Chandran and Patwardhan, 2017). W. somnifera formulation (supplemented with minerals) has been reported to improve both cellular and humoral immunity as well as hematological profile in addition to the significant inhibition in mouse splenocytes (Trivedi et al., 2017). Aqueous root extract of W. somnifera attenuates production of pro-inflammatory cytokines and transcription factor in collagen-induced arthritis (Khan et al., 2018). A study in 2018 showed that W. somnifera significantly inhibited mRNA expression of inflammatory cytokines and promotes the mRNA expression of the anti-inflammatory cytokine in HaCaT cells (Sikandan et al., 2018).

3.4.11 Zingiber officinale Roscoe (Ginger)

Fresh ginger aqueous extract showed antiviral activity against human respiratory syncytial virus in human respiratory tract cell lines (HEp-2 and A549) and decreased the plaque counts in a dose-dependent manner. It also stimulated the secretion of IFN-β that contributes to counteracting against viral infection (Chang et al., 2013). It also showed antiviral potential against avian influenza virus H9N2 on Vero cells in a dose-dependent manner (Rasool et al., 2017). Oral administration of Soft gel capsules containing a Z. officinale in combination showed immunomodulatory and anti-inflammatory properties parallel to those exerted by positive control, and gene expression data highlighted overall same transcriptional remodeling (Dall’Acqua et al., 2019). A study on essential oil of ginger reported immunomodulatory effects by improving the humoral immunity in cyclophosphamide-immunosuppressed mice in a dose-dependent manner (Carrasco et al., 2009). Oral administration of alcoholic ginger extract to allergic rhinitis patients showed significant reduction in total nasal symptom scores (TNSS), with overall improvement in rhino conjunctivitis quality of life questionnaire (Yamprasert et al., 2020). The aqueous and alcoholic extracts of rhizome decreased goblet cell hyperplasia, infiltration of inflammatory cells in airways with reduced total and differential counts of eosinophils and neutrophils in mouse model (Khan et al., 2015) (Table 5).

3.5 Routinely Used Indian Natural Health Supplements to Explore for Use Against COVID 19

3.5.1 Chyawanprash

Chyawanprash is an Ayurvedic polyherbal health supplement, which is made up of concentrated extracts of nutrient-rich herbs and minerals. Chyawanprash comes under Awaleha (electuaries/herbal jams) due to its consistency, and composed of Amla fruit as a base, which is considered as the most active Rasayana to improve strength, stamina, and vitality.

Although several types of research have been published on Chyawanprash to report its health benefits against various ailments, the study reports antioxidant (Anil and Suresh, 2011) free radical scavenging (Bhattacharya et al., 2002) antibacterial, antiviral, anti-inflammatory, antiallergic, and antithrombotic effects (Gupta et al., 2017). In a randomized controlled trial, it was found effective for pulmonary tuberculosis as an adjunct to antitubercular drugs. (Debnath et al., 2012; Sharma R. et al., 2019). An experimental study showed that Chyawanprash pre-treatment reduced plasma histamine levels and IgE release when rats and mice were challenged with allergen- and ovalbumin-induced allergy, suggesting its anti-allergic potential. NK cell activity was significantly increased by Chyawanprash treatment. On treating dendritic cells with Chyawanprash, there was a significant increase in immunity marker levels as well as phagocytic activity that proves its immunomodulatory activity (Sastry et al., 2011).

3.5.2 Triphala

Triphala is a well-known polyherbal Ayurvedic medicine consisting of equal proportions of fruits of Phyllanthus emblica L., Terminalia bellerica (Gaertn.) Roxb. and Terminalia chebula Retz. in the form of powder for digestive and refreshing action. Triphala is associated with many of the therapeutic potentials such as antioxidants, antiinflammatory, antineoplastic, antimicrobial, antidiabetic, etc. (Peterson et al., 2017). Alcoholic extract of Triphala showed specific antimicrobial activity (Tambekar and Dahikar, 2011), broad-spectrum antimicrobial activity against antibiotic-resistant bacteria isolated from humans (Peterson et al., 2017).

Triphala extract was found more active than the NSAID drug, indomethacin, in improving arthritic and inflammatory effects and reduced expression of inflammatory mediators through inhibition of NF-κB activation (Kalaiselvan and Rasool, 2015). In LPS-stimulated macrophages, Triphala inhibited the production of inflammatory mediators, intracellular free radicals, and inflammatory enzymes (Reddy et al., 2009; Kalaiselvan and Rasool, 2016). It has been shown to reduce multiple cell signaling pathways of inflammation and oxidative stress and prevented the noise-stress induced changes in rats thereby strengthening the cell-mediated immune response (Prasad and Srivastava, 2020). A clinical study of Triphala showed immunostimulatory properties on T cells and NK cells, however did not change the cytokine levels in healthy volunteers (Phetkate et al., 2012). The individual constituents of Triphala have also showed immunomodulatory activity (Aher and Wahi, 2011). The stated data on Triphala reveals that it is a powerful polyherbal formulation with countless therapeutic uses for maintaining homeostasis as well as the cure and management of various disease.

3.5.3 Sharbat Rooh Afza

Rooh Afza is a well-known refreshing formulation with global acceptance. It is a concentrated squash prepared as sugar syrup with distillates of numerous medicinal plants including seeds of khurfa (Portulaca oleracea L.), kasni (Cichorium intybus L.), angoor (Vitis vinifera L.), nilofar (Nymphaea alba L.), Neel Kamal (Nymphaea nouchali Burm. f.), kamal (Nelumbo nucifera Gaertn.), gaozaban (Borago officinalis L.), badiyan (Coriandrum sativum L.), fruits/juices of santara (Citrus × sinensis (L.) Osbeck), ananas (Ananas comosus (L.) Merr.), seb (Malus domestica (Suckow) Borkh.), berries (Rubus fruticosus L.), vegetables like palak (Spinacia oleracea L.), gazar (Daucus carota L.), and pudina (Mentha arvensis L.). Rooh Afza boosts the energy system of the body by naturally refreshing. Although there is no evidence on Rooh Afza revealing its therapeutic value, its constituents have been reported as potently antiviral, immunomodulatory, and antiallergic against respiratory disorders.

The flower extract of P. oleracea possessed significant antioxidant and protective effects against DNA damage induced by necrotic effects (Dogan and Anuk, 2019). V. vinifera fruits exhibit anti-asthmatic activity by inhibiting cellular response and subsequent production of inflammatory cytokines (Arora et al., 2016). A study on N. alba flower has been reported against inflammatory activity in Swiss Albino mice using acute inflammatory models in a dose-dependent manners (RS et al., 2013). The immunoregulatory and anti-HIV-1 enzyme activities of N. nucifera suggest that it could be potentially important against virus development (Jiang et al., 2011).

Thus, it can be perceived that Rooh Afza not only provides natural refreshness to the body but also has antioxidant, immunomodulatory, and anti-inflammatory/antiviral activities. However, to validate the scientific data on the therapeutic value of Rooh Afza, experimental research should be undertaken to prove its role in health benefits therapeutically.

The above studies encourage further investigations of traditional medicinal plants for their preventive use against coronavirus infection. The herbs could be taken individually or synergistically at appropriate concentrations as candidates for developing potential therapeutic tools against COVID-19.

3.6 Potential Indian Medicinal Plants for Exploring Against COVID-19

There are many other Indian medicinal plants, which are either part of AYUSH recommendations as such or as ingredients of formulations or are known for improving immunity with antiviral and anti-allergic/anti-inflammatory potential and can offer potential leads against COVID-19. Table 5 provides a list of 83 medicinal plants categorized on a priority basis as per their reported properties. Category 1 (C1) includes 21 “Most promising drugs” which have already shown activity against Coronaviruses/HIV/Dengue viruses with their immunomodulatory and anti-allergic/anti-inflammatory properties. Category 2 (C2) is composed of 44 “Equally promising drugs” which reportedly have shown anti-viral, immunomodulatory, and anti-allergic/anti-inflammatory activities. Category 3 (C3) represents 18 “Possibly promising drugs” which have been reported to show anti-viral/immunomodulatory and/or anti-allergic/anti-inflammatory activities.

Listed medicinal plants and AYUSH recommended formulations could help as the potential alternate therapeutics for management and cure of COVID-19. However, this needs scientific explorations and validation of their preclinical and clinical studies. Since there is such a rich diversity, many other medicinal plants and their bioactive fractions need the attention of the scientific community to be explored against COVID-19.

4 Conclusion

The SARS-CoV-2 has become a threat to human population due to non-availability of approved vaccines or drugs for its treatment. Many herbs that have been reported to work as an immunity booster against other viral infections, and to possess anti-allergic/anti-inflammatory activities, need to be tested against COVID-19. Indian Traditional Medicines have a wide potential for being used in these tough times either for prophylaxis or as adjuvant, owing to their longstanding use in community, ancient references and scientific evidence about their safety and clinical efficacy. The AYUSH ministry, Govt of India has issued several advisories from time to time, considering the strength and evidence of these systems of medicines and making considerable efforts to encourage researchers to explore herbal products for COVID-19. Interventions and herbal formulations from different AYUSH systems have the support of evidence for their immunity-enhancing, anti-inflammatory and antiviral effects. These herbal remedies may, therefore, provide some respite until the availability of trial-tested drug or vaccine to combat the COVID-19 menace. Further, it was noted that a major portion of public and private funding were dedicated to AYUSH trials. More than 50% of these trials were sponsored by the government and various stakeholders associated with the Ministry of AYUSH. It is expected that the results of these clinical studies will be disseminated soon at the public platform so that the policymakers from the AYUSH systems of medicines may reframe their policies for public health and provide information to the global scientific community, which could form a platform for collaborative studies at the national and global levels. The medicinal plant species discussed in this review and categorized for their preclinical and clinical investigation may be taken up by research organizations on priority basis, as this may result in the development of lead molecule against SARS-CoV-2 and COVID-19. Keeping in view the potential of AYUSH medicines and medicinal plants of India, the herbal drug, manufacturers, and the national and global research organizations should develop necessary strategies for furtherance of preclinical and clinical research on these promising therapeutic leads.

Author Contributions

SA: conceptualization, methodology, writing - reviewing and editing; SZ and BP: data curation, writing - original draft preparation; PB, GG, and AP: visualization, investigation; RP and MA: software, validation.

Conflict of Interest

The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Acknowledgments

The corresponding author is thankful to Ministry of AYUSH, Govt of India and DBT for providing funding at Bioactive Natural Product Laboratory, Jamia Hamdard for Research on AYUSH drugs and Medicinal Plants of India.

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Keywords: COVID-19, AYUSH medicine, indian medicinal plants, indian traditional medicine, immunomodulators, antiviral agents

Citation: Ahmad S, Zahiruddin S, Parveen B, Basist P, Parveen A, Gaurav , Parveen R and Ahmad M (2021) Indian Medicinal Plants and Formulations and Their Potential Against COVID-19–Preclinical and Clinical Research. Front. Pharmacol. 11:578970. doi: 10.3389/fphar.2020.578970

Received: 01 July 2020; Accepted: 29 December 2020;
Published: 02 March 2021.

Edited by:

Michael Heinrich, UCL School of Pharmacy, United Kingdom

Reviewed by:

Shailendra Shivaji Gurav, Goa College of Pharmacy, India
Subhash Chandra Mandal, Directorate of Drugs Control, India

Copyright © 2021 Ahmad, Zahiruddin, Parveen, Basist, Parveen, Gaurav, Parveen and Ahmad. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

*Correspondence: Sayeed Ahmad, sahmad_jh@yahoo.co.in

These authors have contributed equally to this work