molecules Communication D8(14)-Ergostenol Glycoside Derivatives Inhibit the Expression of Inflammatory Mediators and Matrix Metalloproteinase Hyejin Moon 1,†, Myoungsil Ko 1,†, Yujin Park 2,†, Jeonguk Kim 1, Dowon Yoon 1, Eunjoohwang Lee 2, Taehoon Lee 1,* and Hakwon Kim 1,* 1 Global Center for Pharmaceutical Ingredient Materials, Department of Applied Chemistry, Kyung Hee University, Yongin 17104, Korea; [email protected] (H.M.); [email protected] (M.K.); [email protected] (J.K.); [email protected] (D.Y.) 2 Graduate School of East-West Medicinal Science, Kyung Hee University, Yongin 17104, Korea; [email protected] (Y.P.); [email protected] (E.L.) * Correspondence: [email protected] (T.L.); [email protected] (H.K.); Tel.: +823-1201-5317 (T.L.); +823-1201-2459 (H.K.) † These authors equally contribute to this work. Abstract: Arthritis is a chronic inflammatory disease accompanied by pathological reactions such as swelling, redness, fever, and pain in various joint areas. The drugs currently available to treat arthritis are associated with diverse side-effects. Therefore, there is a need for safer and more effective treatments to alleviate the inflammation of arthritis with fewer side-effects. In this study, a new sterol, D8(14)-ergostenol, was discovered, and its glycosides were synthesized and found to be more efficient in terms of synthesis or anti-inflammatory activity than either spinasterol or 5,6-dihydroergosterol is. Among these synthetic glycosides, galactosyl ergostenol inhibited the expression of inflammatory mediators in TNF-α-stimulated FLS and TNF-α-induced MMPs and collagen type II A1 degradation Citation: Moon, H.; Ko, M.; Park, Y.; in human chondrocytes. These results suggest the new galactosyl ergostenol as a treatment candidate Kim, J.; Yoon, D.; Lee, E.; Lee, T.; for arthritis. Kim, H. D8(14)-Ergostenol Glycoside Derivatives Inhibit the Expression of Keywords: ergostenol; ergostenol glycosides; arthritis; matrix metalloproteinase; collagen type II A1 Inflammatory Mediators and Matrix Metalloproteinase. Molecules 2021, 26, 4547. https://doi.org/10.3390/ molecules26154547 1. Introduction Arthritis is a chronic inflammatory disease accompanied by pathological reactions Academic Editor: Michael John Plater such as swelling, redness, fever, and pain in various joint areas [1–4]. Joints are areas where two or more bones meet, and most of them are responsible for helping the body move [2]. Received: 1 July 2021 In terms of anatomy, the structure of a joint is composed of cartilage, which protects the Accepted: 23 July 2021 Published: 28 July 2021 end of the bone, and a synovial membrane that secretes a clear sticky fluid around the joint [3]. Joint tissue inflammation is classified as either degenerative osteoarthritis (OA) or Publisher’s Note: MDPI stays neutral rheumatoid arthritis (RA) according to the loss of cartilage and autoimmune dysfunction with regard to jurisdictional claims in of the synovial membrane [4]. published maps and institutional affil- The major cause of OA is age-related damage to chondrocytes. The primary patho- iations. logical mechanism of OA is the reduced formation of the extracellular matrix of articular cartilage cells and the alteration of the extracellular matrix components of cartilage due to cellular aging [5]. Normal chondrocytes play an important role in cartilage homeostasis. Abnormal chondrocytes exposed to inflammatory cytokines and chemokines cause the excessive secretion of several types of matrix-degrading enzymes such as matrix metal- Copyright: © 2021 by the authors. Licensee MDPI, Basel, Switzerland. loproteinases (MMPs). These MMPs break down collagen type II A1 (COL2A1), a major This article is an open access article collagen component of the cartilage matrix. In particular, MMP-1, MMP-3, and MMP-13 distributed under the terms and play a major role in arthritis [6–8]. Rheumatoid arthritis is a chronic inflammatory condi- conditions of the Creative Commons tion that arises in the synovial membrane surrounding the joints. The major pathogenic Attribution (CC BY) license (https:// mechanism of RA is the excessive secretion of inflammatory cytokines and chemokines creativecommons.org/licenses/by/ due to immune abnormalities, leading to the gradual destruction of cartilage tissue with an 4.0/). increased production of matrix degradation enzymes such as MMPs [9]. Molecules 2021, 26, 4547. https://doi.org/10.3390/molecules26154547 https://www.mdpi.com/journal/molecules Molecules 2021, 26, x 2 of 12 Molecules 2021, 26, 4547 due to immune abnormalities, leading to the gradual destruction of cartilage tissue2 with of 12 an increased production of matrix degradation enzymes such as MMPs [9]. The major constituents of the cartilage matrix are COL2A1 and proteoglycans. De- generative osteoarthritis and rheumatoid arthritis are caused by the degradation of COL2A1,The major which constituents is primarily of mediated the cartilage by matrixMMP-1, are MMP-3, COL2A1 and and MMP-13. proteoglycans. These Degen-MMPs erative osteoarthritis and rheumatoid arthritis are caused by the degradation of COL2A1, are secreted mainly by human chondrocytes (HCs) and human fibroblast-like synovio- which is primarily mediated by MMP-1, MMP-3, and MMP-13. These MMPs are secreted cytes (FLSs) [4]. The over-expression of MMPs such as MMP-1, MMP-3, and MMP-13 mainly by human chondrocytes (HCs) and human fibroblast-like synoviocytes (FLSs) [4]. leads to a vicious cycle of the activation of inflammatory cytokines and chemokines [10]. The over-expression of MMPs such as MMP-1, MMP-3, and MMP-13 leads to a vicious cycle Regulating these inflammatory mediators can alleviate symptoms of arthritis. Currently, of the activation of inflammatory cytokines and chemokines [10]. Regulating these inflam- various drugs, including steroidal, nonsteroidal, and biological materials with analgesic matory mediators can alleviate symptoms of arthritis. Currently, various drugs, including or anti-inflammatory properties, are used to treat arthritis [11]. These drugs generally steroidal, nonsteroidal, and biological materials with analgesic or anti-inflammatory prop- have a short duration of effect and low efficacy, and they can cause many side effects erties, are used to treat arthritis [11]. These drugs generally have a short duration of when used long term [12,13]. Thus, there is a need for safer and more effective arthritis effect and low efficacy, and they can cause many side effects when used long term [12,13]. treatments to alleviate joint inflammation and inhibit COL2A1 degradation through inhi- Thus, there is a need for safer and more effective arthritis treatments to alleviate joint inflammationbition of MMPs. and inhibit COL2A1 degradation through inhibition of MMPs. Previously,Previously, we we reported reported the the potent potent anti-inflammatory anti-inflammatory activity activity of of 5,6-dihydroergosterol 5,6-dihydroergoste- glucoserol glucose (DHE-Glu, (DHE-Glu,2a), 2a a), synthetic a synthetic compound compound derived derived from from a a natural natural product,product, spinas-spinas- terolterol glucoseglucose (Spin-Glu, 1a1a))[ [14].14]. In In HaCaT HaCaT cells, cells, DHE-Glu DHE-Glu (2a (2a) produced) produced strong strong anti-in- anti- inflammatoryflammatory activity activity through through the the inhibition ofof TNF-TNF-αα/IFN-/IFN-γ-induced-induced expressionsexpressions ofof CCL17CCL17 andand CCL22,CCL22, andand alsoalso inhibitedinhibited DNCB-inducedDNCB-induced contactcontact dermatitisdermatitis inin anan animalanimal modelmodel of chronic chronic inflammation inflammation [15]. [15]. As As a follow-up a follow-up to this to this study, study, we wetried tried to find to finda new a newsynthetic synthetic sterol sterolthat could that be could synthesized be synthesized more efficiently more efficiently and/or had and/or a greater had bioactivity a greater bioactivitythan spinasterol than spinasterol(Spin, 1) or (Spin, 5,6-dihydroergosterol1) or 5,6-dihydroergosterol (DHE, 2) (DHE,did. As2 )a did. result, As aΔ result,8(14)-er- Dgostenol8(14)-ergostenol (Ergn, 3 (Ergn,), a sterol3), ahaving sterol havinga double a doublebond at bond carbon at carbonposition position 8(14) of 8(14) the steroid of the steroidskeleton, skeleton, was identified was identified as a candidate as a candidate compound. compound. This sterol This sterolcan be can synthesized be synthesized from fromergosterol ergosterol by selective by selective hydrogenation hydrogenation followed followed by double by double bond bond migration migration [16]. [We16]. syn- We synthesizedthesized ergostenol ergostenol glycosides glycosides containing containing a asu sugar,gar, such such as as glucose, glucose, galactose, galactose, or xylose, byby SchmidtSchmidt glycosylationglycosylation ofof ErgnErgn ((33)) withwith aa glycosylglycosyl imidate.imidate. SpinSpin ((11),), Spin-GluSpin-Glu ((1a1a),), DHEDHE ((22),), DHE-GluDHE-Glu ((2a2a),), ErgnErgn ((33),), andand ergostenolergostenol glycosidesglycosides includingincluding Ergn-GluErgn-Glu ((3a3a),), Ergn-GalErgn-Gal ((3b3b),), andand Ergn-XylErgn-Xyl ((3c3c)) areare shownshown inin FigureFigure1 1.. In In this this study, study, newly newly synthesized synthesized ergostenol ergostenol glycosidesglycosides werewere testedtested forfor anti-inflammatoryanti-inflammatory propertiesproperties inin FLSsFLSs andand thethe suppressionsuppression ofof MMPsMMPs expressionexpression inin HCs. HCs. These These results results may may suggest suggest the the possibility possibility of beingof being